ABSTRACT
Target-based high-throughput screening (HTS) plays an integral role in drug discovery. The implementation of HTS assays generally requires high expression levels of the target protein, and this is typically accomplished using recombinant cDNA methodologies. However, the isolated gene sequences to many drug targets have intellectual property claims that restrict the ability to implement drug discovery programs. The present study describes the pharmacological characterization of the human histamine H3 receptor that was expressed using random activation of gene expression (RAGE), a technology that over-expresses proteins by up-regulating endogenous genes rather than introducing cDNA expression vectors into the cell. Saturation binding analysis using [125I]iodoproxyfan and RAGE-H3 membranes revealed a single class of binding sites with a K(D) value of 0.77 nM and a B(max) equal to 756 fmol/mg of protein. Competition binding studies showed that the rank order of potency for H3 agonists was N(alpha)-methylhistamine approximately (R)-alpha- methylhistamine > histamine and that the rank order of potency for H3 antagonists was clobenpropit > iodophenpropit > thioperamide. The same rank order of potency for H3 agonists and antagonists was observed in the functional assays as in the binding assays. The Fluorometic Imaging Plate Reader assays in RAGE-H3 cells gave high Z' values for agonist and antagonist screening, respectively. These results reveal that the human H3 receptor expressed with the RAGE technology is pharmacologically comparable to that expressed through recombinant methods. Moreover, the level of expression of the H3 receptor in the RAGE-H3 cells is suitable for HTS and secondary assays.
Subject(s)
Gene Expression/drug effects , Receptors, Histamine H3/genetics , Transfection/methods , Binding, Competitive , Cell Line, Tumor , Fluorometry/methods , Genetic Vectors/genetics , Histamine Agonists/pharmacology , Histamine Antagonists/pharmacology , Humans , Imidazoles/metabolism , Imidazoles/pharmacology , Iodine Radioisotopes , Radioligand Assay , Receptors, Histamine H3/metabolism , Technology, Pharmaceutical/methodsABSTRACT
The serotonin (5-hydroxytryptamine) 5-HT2 receptor subfamily consists of three members, 5-HT2A, 5-HT2B, and 5-HT2C. These receptors share high homology in their amino acid sequence, have similar signaling pathways, and have been indicated to play important roles in feeding, anxiety, aggression, sexual behavior, mood, and pain. Subtype-selective agonists and antagonists have been explored as drugs for hypertension, Parkinson's disease, sleep disorders, anxiety, depression, schizophrenia, and obesity. In this study, we report the development of homogeneous agonist binding assays in a scintillation proximity assay (SPA) format to determine the high-affinity binding state of agonist compounds for the human 5-HT2C, 5-HT2A, and 5-HT2B receptors. The 5-HT2 agonist 1-(4- [125I]iodo-2,5-dimethoxyphenyl)-2-aminopropane ([125I]DOI) was used to label the high-affinity sites for the 5-HT2A and 5-HT2C receptors. The high-affinity sites for the 5-HT2B receptor were labeled with [3H]lysergic acid diethylamide. Total receptor expression was determined with the 5-HT2 antagonist [3H]mesulergine for the 5-HT2B and 5-HT2C receptors, and [3H]ketanserin for the 5-HT2A receptor. The agonist high-affinity binding sites accounted for 2.3% (5-HT(2C) receptor), 4.0% (5-HT2A receptor), and 22% (5-HT2B receptor) of the total receptor population. Competition binding studies using known agonists indicated high Z' values of the agonist binding assays in SPA format (Z' > 0.70). The Ki values of 5-HT, (R)(-)DOI, and VER-3323 for the 5-HT2A, 5-HT2B, and 5-HT2C receptors by SPA format were equivalent to published data determined by filtration binding assays. These results indicate that agonist binding assays in SPA format can be easily adapted to a high throughput assay to screen for selective 5-HT2C receptor agonists, as well as for selectivity profiling of the compounds.
Subject(s)
Drug Evaluation, Preclinical/methods , Serotonin 5-HT2 Receptor Agonists , Serotonin Receptor Agonists/pharmacology , Amphetamines/pharmacology , Binding, Competitive , Calcium Signaling/drug effects , Cell Line , Dose-Response Relationship, Drug , Ergolines/metabolism , Humans , Ketanserin/metabolism , Lysergic Acid Diethylamide/pharmacology , Radioligand Assay , Receptor, Serotonin, 5-HT2A/analysis , Receptor, Serotonin, 5-HT2A/metabolism , Receptor, Serotonin, 5-HT2B/analysis , Receptor, Serotonin, 5-HT2B/metabolism , Receptor, Serotonin, 5-HT2C/analysis , Receptor, Serotonin, 5-HT2C/metabolism , Serotonin/pharmacology , Serotonin Antagonists/metabolism , TransfectionABSTRACT
BACKGROUND: Ghrelin, an endogenous ligand for growth hormone secretagogue receptor (GHS-R), is known to increase food intake in lean humans and rodents. In addition, ghrelin levels are increased by fasting in lean rodents and are elevated before meals in humans, suggesting an important role for ghrelin in meal initiation. However, in obese human, circulating ghrelin levels were found to be significantly reduced as compared to lean individuals. OBJECTIVES: To evaluate whether circulating ghrelin levels, as well as ghrelin sensitivity, are decreased in obese individuals in order to limit its effect on food intake. DESIGN: : Lean C57BL/6J mice fed a chow, a low- (LFD) or a high-fat diet (HFD) were used to determine ghrelin regulation and secretion as well as ghrelin sensitivity. MEASUREMENTS: Plasma ghrelin levels were measured in low- and high-fat fed mice. Ghrelin-induced food intake was measured in chow, low- and high-fat fed mice. RESULTS: We measured ghrelin levels in lean and diet-induced obese mice, fed on an LFD or an HFD, respectively. We observed that not only ghrelin secretion was reduced in obese mice but its diurnal regulation was also lost. In addition, we failed to observe any change in ghrelin secretion upon fasting and refeeding. Moreover, we observed that the sensitivity to the orexigenic effects of exogenous ghrelin was reduced in obese mice when compared to lean mice fed a chow or a LFD. The insensitivity of obese mice to ghrelin was improved upon weigh loss. CONCLUSION: : Altogether, these results indicate that ghrelin secretion and regulation is impaired in dietary-induced obesity in mice and suggest that ghrelin inhibition could prevent weight regain after weight loss.
Subject(s)
Obesity/blood , Peptide Hormones/blood , Weight Loss , Animals , Circadian Rhythm , Dietary Fats/administration & dosage , Dose-Response Relationship, Drug , Eating/drug effects , Fasting/blood , Ghrelin , Mice , Mice, Inbred C57BL , Obesity/physiopathology , Peptide Hormones/metabolism , Peptide Hormones/pharmacology , Weight Loss/physiologyABSTRACT
Monosodium-glutamate (MSG) is neurotoxic for brain regions devoid of blood-brain barrier when it is injected at high doses during the neonatal period. Neuropeptide Y (NPY) neurons in the arcuate nucleus are particularly sensitive to MSG treatment. But, despite of the large decrease of this potent orexigenic peptide, feeding behavior is only slightly affected. We hypothesized that the hypothalamic NPY receptor system might be modified in these rats. The present study characterizes hypothalamic NPY and NPY receptors in normal and MSG-treated rats. MSG-treated rats were lighter (p < 0.01) and ate 17% less than the control rats (p < 0.01). NPY levels in the mediobasal and mediodorsal hypothalamus were reduced in MSG-treated rats compared to normal rats (-26% and -43%, p < 0.05 and p < 0.01, respectively). Combined hypothalamic Y1 and Y5 NPY receptor density was increased in MSG-treated rats compared to normal rats (+25%, p < 0.04), but affinity remained unaltered. Blockade with a selective Y1 antagonist showed that the Y1 receptor subtype represented more than 90% of the combined Y1 and Y5 receptor populations. The up-regulation of the NPY receptors is an element necessary to maintain food intake at a sufficient level to allow survival and growth of the lesioned rats.
Subject(s)
Arginine/analogs & derivatives , Hypothalamus/chemistry , Hypothalamus/drug effects , Receptors, Neuropeptide Y/analysis , Sodium Glutamate/pharmacology , Animals , Arginine/pharmacology , Body Weight , Eating , Neuropeptide Y/analysis , Rats , Rats, Sprague-Dawley , Receptors, Neuropeptide Y/antagonists & inhibitorsABSTRACT
OBJECTIVE: Leptin plays a major role in the regulation of body weight. It circulates in both free and bound form. One of the leptin receptor isoforms exists in a circulating soluble form that can bind leptin. In the present study, we measured the soluble leptin receptor (SLR) levels in lean and obese humans. We investigated the relationship between plasma SLR levels, plasma leptin levels and the degree of obesity. We also examined whether SLR concentrations could be modulated by fat mass loss induced by a 3 month weight-reducing diet. SUBJECTS: A total of 112 obese (age 18-50 y; body mass index (BMI) 30-44 kg/m2; 23 men and 89 women), 38 overweight (age 19-48 y; BMI 25-29 kg/m2; 10 men and 28 women) and 63 lean (age 18-50 y; BMI 17-24 kg/m2; 16 men and 47 women) humans. MEASUREMENTS: A direct double monoclonal sandwich enzyme-linked immunosorbent assay (ELISA) was used for the quantitative measurement of the soluble human leptin receptor. Leptin was measured by radioimmunoassay (RIA). Body composition was assessed by biphotonic absorptiometry DEXA (dual energy X-ray absorptiometry). RESULTS: We observed that the SLR is present in human plasma (range 10-100 ng/ml). SLR levels were lower in obese and overweight than lean subjects (28.7+/-8.8, 40.2+/-14.9, 51.2+/-12.5 ng/ml, respectively) and were inversely correlated to leptin and percentage of body fat (r=-0.74 and r=-0.76; respectively; P<0.0001). The ratio of circulating leptin to SLR was strongly related to the percentage of body fat (r=0.91; P<0.0001). Interestingly a gender difference was observed in SLR levels, which were higher in obese and overweight men than in obese and overweight women. In obese subjects after a 3 month low-calorie diet, SLR levels increased in proportion to the decrease in fat mass. In the gel filtration profile, SLR coeluted exactly with the bound leptin fractions. CONCLUSION: Obesity, in humans is associated with decreasing levels of the circulating soluble leptin receptor (SLR). The relationship of SLR with the degree of adiposity suggests that high SLR levels may enhance leptin action in lean subjects more than in obese subjects.
Subject(s)
Carrier Proteins/blood , Obesity/blood , Receptors, Cell Surface , Adipose Tissue , Adult , Body Composition , Body Mass Index , Chromatography, Gel , Diet, Reducing , Energy Intake , Enzyme-Linked Immunosorbent Assay , Female , Humans , Leptin/blood , Male , Middle Aged , Radioimmunoassay , Receptors, Leptin , Sex CharacteristicsABSTRACT
Many hyothalamic neuropeptides are involved in the regulation of food intake and body weight. The orexins (OX) which are synthesized in the lateral hypothalamus are among the most recently characterized whereas neuropeptide Y (NPY) belongs to a group of "older" peptides extensively studied for their effects on feeding behavior. Both stimulate food ingestion in rodents. In this experiment, we measured the expressions of these peptides as well as of their receptors (OX1-R and OX2-R, Y1 and Y5) in the hypothalamus of obese hyperphagic and lean Zucker rats by real-time RT-PCR using the TaqMan apparatus. NPY mRNA expression in the obese rats was significantly increased by a factor of 10 (P < 0.002) whereas expressions of the Y1 and Y5 receptors were decreased by 25% (P < 0.01) and 50% (P < 0.002), respectively. Their prepro-orexin mRNA expression was more than twofold decreased (P < 0.01) and expressions of their OX receptors 1 and 2 mRNA were five- and fourfold increased (P < 0.05), respectively. An inverse phenomenon was therefore noted between the two peptides: for NPY, increased levels and downregulation of receptors; and for OX, diminished levels with upregulation of receptors. The reasons for these changes might be linked to the absence of leptin signaling as similar profiles are found in the ob/ob mice. For orexins at least, other factors such as hyperglycemia might be involved. Based on anatomical considerations, a direct effect of NPY or of other brain peptides such as CRH cannot be excluded. We conclude that the diminution in the OX tone might participate in a counterregulatory system necessary to limit the noxious effects of NPY on food intake and body weight.
Subject(s)
Carrier Proteins/biosynthesis , Hypothalamus/physiology , Intracellular Signaling Peptides and Proteins , Neuropeptide Y/biosynthesis , Neuropeptides/biosynthesis , Obesity/metabolism , Receptors, Neuropeptide/biosynthesis , Animals , Body Weight , Carrier Proteins/genetics , Eating , Leptin/blood , Male , Neuropeptide Y/genetics , Neuropeptides/genetics , Orexin Receptors , Orexins , Peptides/genetics , Peptides/metabolism , Protein Precursors/biosynthesis , Protein Precursors/genetics , RNA, Messenger/biosynthesis , Rats , Rats, Zucker , Receptors, G-Protein-Coupled , Receptors, Neuropeptide/genetics , Receptors, Neuropeptide Y/biosynthesis , Receptors, Neuropeptide Y/geneticsABSTRACT
Melanin concentrating hormone (MCH) is a peptide synthesized in the lateral hypothalamus which stimulates food ingestion and leptin secretion in rodents. In this experiment, we measured the expressions of MCH as well as of its receptor (SLC-1) in the hypothalamus of obese hyperphagic and lean Zucker rats by quantitative real time RT-PCR. MCH mRNA expression in the obese rats was significantly increased by a factor of five (P<0.01) whereas expression of SLC-1 was decreased by more than 50% (P<0.05). Circulating levels of leptin and MCH were increased in the plasma of obese Zucker rats when compared to lean rats (38-fold and 1.7-fold, respectively, P<0.001 and P<0.01). However, individual MCH levels were not directly correlated to leptin levels in the lean (functional leptin receptor) or in the obese (non-functional leptin receptor) Zucker rats. These results indicate that the absence of leptin signaling in rats is associated with an increased hypothalamic expression and circulating release of MCH, contributing to their obesity syndrome.
Subject(s)
Hypothalamic Area, Lateral/metabolism , Hypothalamic Hormones/metabolism , Melanins/metabolism , Obesity/metabolism , Pituitary Hormones/metabolism , Receptors, Cell Surface , Animals , Carrier Proteins/genetics , Disease Models, Animal , Gene Expression Profiling , Genotype , Leptin/blood , Leptin/metabolism , Male , Obesity/genetics , Point Mutation , Protein Precursors/metabolism , Rats , Rats, Zucker , Receptors, Leptin , Receptors, Pituitary Hormone/metabolismABSTRACT
OBJECTIVES: To examine whether the survival (both overall and disease-specific) of patients who underwent radical prostatectomy varies from region to region in the United States. Previous reports have documented a geographic variation in the use of radical prostatectomy. METHODS: This study was based on the data from nine geographic regions of the Surveillance, Epidemiology, and End Results Program (SEER) for 1983 through 1992. Patients with localized prostate cancer who underwent radical prostatectomy were included in the analysis. A proportional hazards model was used to investigate whether geographic variation is associated with both overall and disease-specific survival. RESULTS: From 1983 through 1992, the SEER Program collected information from nine geographic regions on 66,293 patients with localized prostate cancer (mean age 71.8 +/- 8.4 years), who had SEER grade codes of 1, 2, or 3. Of these patients, 11,429 (mean age 65.3 +/- 6.5 years) underwent radical prostatectomy and lymph node dissection. Cox's proportional hazards analyses revealed that the impact of geographic location on both overall and disease-specific survival in patients who underwent radical prostatectomy was not statistically significant. CONCLUSIONS: The results of this study indicate that the survival (both overall and disease-specific) of patients with localized prostate cancer who underwent radical prostatectomy is not influenced by geographic location, suggesting that their survival is relatively uniform across the geographic regions in the United States.
Subject(s)
Prostatectomy/statistics & numerical data , Prostatic Neoplasms/mortality , Aged , Analysis of Variance , Disease-Free Survival , Humans , Male , Middle Aged , Proportional Hazards Models , Prostatic Neoplasms/surgery , SEER Program , Survival Rate , United States/epidemiologyABSTRACT
OBJECTIVES: To examine the geographic variation in the use of radiation to treat nonmetastatic prostate cancer and to identify factors that explain the variation in the United States. METHODS: This study was based on data from the nine geographic regions of the Surveillance, Epidemiology, and End Results Program for 1983 through 1996. Patients with localized or regional prostate cancer who did not undergo surgical treatment were included in the analysis. Logistic regression analysis was used to investigate the influence of geographic and demographic factors on the use of radiation. The squared multiple correlation coefficient R(2) was used to measure the proportion of variation in the selection of radiation explained by each factor of interest. RESULTS: Compared with San Francisco, the adjusted odds ratios for 6 of the 8 geographic areas had highly significant P values, suggesting the use of radiation therapy varies from region to region. However, geographic location only explained less than 3% of the total variation in the use of radiation. The geographic location explained a much higher proportion of variation in the youngest (younger than 55 years) and the oldest (80 years old or older) groups. Overall, age was the most important factor that influenced the use of radiation. CONCLUSIONS: The finding that geographic location explains a significant proportion of the variation in the use of radiation in the youngest and oldest age groups demonstrates the outcome of longstanding controversies in the nonsurgical treatment of prostate cancer. Documenting the impact of the interaction of age and geographic location on the treatment approaches provides for better understanding of the impact of patients and physicians making clinical decisions in the management of nonmetastatic prostate cancer.
Subject(s)
Prostatic Neoplasms/radiotherapy , Black or African American/statistics & numerical data , Age Factors , Aged , Aged, 80 and over , Hispanic or Latino/statistics & numerical data , Humans , Male , Middle Aged , Prostatic Neoplasms/ethnology , Regression Analysis , SEER Program , United StatesABSTRACT
Neuropeptides present in the hypothalamus and new messengers in the periphery such as leptin modulate food intake in mammals. Neuropeptide Y (NPY) and galanin in microdissected brain areas and plasma leptin levels were measured by specific radioimmunoassays during the resting period in rats selected for their strong preference either for carbohydrate or fat, but with identical energy intake. NPY concentrations were 23% lower (p <.02) in carbohydrate-preferring (CP) than in fat-preferring (FP) rats in the parvocellular part of the paraventricular nucleus (PVN), which is one of the main areas involved in the regulation of feeding behavior. On the other hand, galanin was significantly (+25%, p = .03) higher in CP rats than in FP rats in the magnocellular part of the PVN. Plasma leptin was more than 50% higher in FP rats than in CP rats (p < .01) and highly correlated with the fat preference (r = 0.57; p = .003) and body weight gain. We conclude that the rats with a spontaneous and marked dietary preference have a characteristic peptidergic profile. Due to their anatomical relationships, neuropeptide Y could act in conjunction with galanin in a peptidergic balance located in the paraventricular nucleus. This model integrates information provided by the energy stores and translated by peripheral messengers such as leptin which could act in a counterregulatory manner in order to limit the overweight induced by the ingestion of unbalanced diets.
Subject(s)
Feeding Behavior/physiology , Food Preferences/physiology , Hypothalamus/metabolism , Leptin/blood , Neuropeptide Y/metabolism , Animals , Dietary Fats , Energy Intake , Energy Metabolism , Galanin/metabolism , Male , Paraventricular Hypothalamic Nucleus/metabolism , Radioimmunoassay , Rats , Rats, Long-Evans , Regression Analysis , Sensitivity and Specificity , Weight GainABSTRACT
Long-form leptin receptor (OB-R(L)) is a signal-transducing member of the cytokine receptor superfamily that is essential for mediating the effects of leptin on mammalian body weight homeostasis. At present, the range of transcriptional targets responsive to OB-R(L) activation, and consequently, the likely mediators of leptin action, remain undefined. In this report, we have used cDNA subtractive hybridization to identify transcripts induced by leptin in immortalized hypothalamic neurons expressing OB-R(L). Differential expression of the identified transcripts in these cells was confirmed by both array technology and Northern blotting. In situ hybridization studies indicate that these transcripts are expressed in the mouse central nervous system, including nuclei of the hypothalamus that coexpress OB-R(L). Comparative in situ analysis of slices of hypothalami generated from control and leptin-injected ob/ob mice demonstrates that a subset of the identified transcripts is induced in vivo after leptin injection. The potential role of the proteins encoded by these transcripts in mediating the effects of leptin on body weight and energy homeostasis are discussed.
Subject(s)
Brain/metabolism , Carrier Proteins/genetics , Hypothalamus/metabolism , Leptin/pharmacology , Neurons/metabolism , Receptors, Cell Surface , Transcription, Genetic/drug effects , Adenoviridae , Animals , Cell Line , Gene Expression Regulation/drug effects , Gene Library , Genetic Vectors , In Vitro Techniques , Male , Mice , Mice, Obese , Organ Specificity , Receptors, Leptin , Recombinant Proteins/biosynthesis , TransfectionSubject(s)
Eating/physiology , Neuropeptide Y/metabolism , Oligodeoxyribonucleotides, Antisense , Receptors, Neuropeptide Y/metabolism , Animals , Cells, Cultured , Galanin/metabolism , Humans , Hypothalamus/metabolism , Mice , RNA, Messenger/metabolism , Radioligand Assay , Rats , Reverse Transcriptase Polymerase Chain Reaction , ThionucleotidesABSTRACT
Protein-tyrosine phosphatase 1B (PTP-1B) is a major protein-tyrosine phosphatase that has been implicated in the regulation of insulin action, as well as in other signal transduction pathways. To investigate the role of PTP-1B in vivo, we generated homozygotic PTP-1B-null mice by targeted gene disruption. PTP-1B-deficient mice have remarkably low adiposity and are protected from diet-induced obesity. Decreased adiposity is due to a marked reduction in fat cell mass without a decrease in adipocyte number. Leanness in PTP-1B-deficient mice is accompanied by increased basal metabolic rate and total energy expenditure, without marked alteration of uncoupling protein mRNA expression. In addition, insulin-stimulated whole-body glucose disposal is enhanced significantly in PTP-1B-deficient animals, as shown by hyperinsulinemic-euglycemic clamp studies. Remarkably, increased insulin sensitivity in PTP-1B-deficient mice is tissue specific, as insulin-stimulated glucose uptake is elevated in skeletal muscle, whereas adipose tissue is unaffected. Our results identify PTP-1B as a major regulator of energy balance, insulin sensitivity, and body fat stores in vivo.
Subject(s)
Adipose Tissue/physiology , Energy Metabolism , Insulin Resistance/genetics , Membrane Transport Proteins , Mitochondrial Proteins , Protein Tyrosine Phosphatases/deficiency , Animals , Body Weight/genetics , Carrier Proteins/genetics , Female , Glucose/metabolism , Glucose Tolerance Test , Homeostasis , Hyperinsulinism/metabolism , Ion Channels , Leptin/genetics , Male , Membrane Proteins/genetics , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Muscle, Skeletal/metabolism , Protein Tyrosine Phosphatase, Non-Receptor Type 1 , Protein Tyrosine Phosphatases/genetics , Proteins/genetics , RNA, Messenger , Uncoupling Protein 1 , Uncoupling Protein 2 , Uncoupling Protein 3ABSTRACT
Serum prostate markers, in particular prostate-specific antigen (PSA), have truly revolutionised all aspects of the management of men with prostatic carcinoma (PCa), the most important application being related to its early detection and screening. Several studies have shown the clinical utility of PSA levels for staging patients with PCa, especially when associated with other parameters, such as tumour grade, digital rectal examination and transrectal ultrasound findings, to establish the likelihood of disease extension outside the gland and of positive lymph nodes. Also, serum PSA levels are useful in monitoring patients either after the initial diagnosis of PCa or following therapy.
Subject(s)
Prostate-Specific Antigen/blood , Prostatic Diseases/blood , Carcinoma/blood , Carcinoma in Situ/blood , Humans , Male , Prostate/metabolism , Prostate/pathology , Prostate-Specific Antigen/metabolism , Prostatic Diseases/therapy , Prostatic Neoplasms/bloodABSTRACT
OBJECTIVES: Previous reports have documented a geographic variation in the use of radical prostatectomy. We examined whether this phenomenon can be explained by factors other than geography alone. METHODS: This study was based on the data from nine geographic regions of the Surveillance, Epidemiology, and End Results (SEER) program for the years 1983 through 1994. Patients with localized or regional prostate cancer were included in the analysis. Logistic regression analysis was used to investigate the influence of geographic and demographic factors on the use of radical prostatectomy. The squared multiple correlation coefficient R(2) was used to measure the proportion of variation in the selection of radical prostatectomy explained by each factor of interest. RESULTS: As previously reported, the use of radical prostatectomy was significantly associated with geographic location; the degree of geographic variation varied as a function of age and was most dramatic in the youngest (younger than 45 years) and the oldest (75 years or older) groups. Overall, however, geography explained less than 2% of the total variation in the use of radical prostatectomy. Age was the most important factor that influenced the use of radical prostatectomy. CONCLUSIONS: Geography explains only a small proportion of the variation in the use of radical prostatectomy. In fact, of the factors examined, only age appeared to meaningfully explain the variation in the use of radical prostatectomy. Overall, our ability to explain the variation in the use of radical prostatectomy remains meager, and new factors must be identified if we are to better understand how patients and physicians make clinical decisions.
Subject(s)
Prostatectomy/statistics & numerical data , SEER Program , Adult , Age Distribution , Aged , Aged, 80 and over , Demography , Humans , Male , Middle Aged , United StatesABSTRACT
The radical prostatectomy has been modified over the years. With the introduction to the modern operating room of robots and other tools has come the latest modification: the laparoscopic radical prostatectomy (LRP), first described almost 10 years ago. In the past 2 years, the technique of LRP has been made standard, reproducible, and efficient. The LRP virtually eliminates the physical and emotional toll of radical prostate surgery and reduces blood loss, hospital time, and cost. Published series demonstrate oncologic and functional results comparable with and perhaps better than what is seen with open radical prostatectomy. The LRP is a standard surgical technique in a growing number of medical centers, with world experience estimated at 500 cases. The LRP offers the urologic community a rare opportunity for tangibly improving care.
Subject(s)
Laparoscopy/methods , Prostatectomy/methods , Prostatic Neoplasms/surgery , Humans , MaleABSTRACT
OBJECTIVE: The purpose of the present study was to investigate the continuing validity of the hypothesis that leptin is a physiologically important regulator of food intake, using the human leptin mutant R128Q leptin. DESIGN: In a cellular proliferation assay, based on BAF-3 cells transfected with the murine ObRb receptor, R128Q leptin was shown to be devoid of agonistic activity and to competitively inhibit the proliferative effects of leptin. To determine whether R128Q leptin was also an antagonist of leptin in vivo, the leptin mutant was injected intracerebroventricularly (i.c.v.) into rats in the absence and presence of leptin. R128Q was also injected intraperitoneally (i.p.) into ob/ob and into db/db mice expressing, respectively, either normal or defective ObRb receptors. RESULTS: R128Q was shown to be a competitive antagonist of leptin induced cellular proliferation in vitro. Surprisingly, in vivo R128Q leptin produced a strong dose-dependent decrease in food intake, and was only slightly less potent than leptin itself. In fasted rats, the inhibitory effects of leptin and R128Q leptin (i.c.v.) on post-fast refeeding were additive. Finally, R128Q leptin produced the same inhibition of food intake as leptin when injected i.p. in ob/ob mice and, like leptin, was inactive after i.p. injection to db/db mice. CONCLUSION: R128Q leptin is a leptin agonist in vivo, but behaves as an antagonist against leptin induced proliferation in vitro. The data demonstrate that the human leptin mutant R128Q leptin is not a suitable tool for investigating the physiological actions of leptin.
Subject(s)
Eating/drug effects , Obesity/metabolism , Proteins/antagonists & inhibitors , Proteins/pharmacology , Receptors, Cell Surface , Animals , Carrier Proteins/metabolism , Cell Division/drug effects , Cell Line/drug effects , Dose-Response Relationship, Drug , Female , Injections, Intraperitoneal , Injections, Intraventricular , Leptin , Male , Mice , Mice, Obese , Proteins/administration & dosage , Proteins/metabolism , Rats , Rats, Sprague-Dawley , Receptors, LeptinABSTRACT
It has been claimed that factors favoring the development or maintenance of animal or human obesity may include increases in glucocorticoid production or hyperresponsiveness of the hypothalamic-pituitary-adrenal axis. In normal rats, glucocorticoids have been shown to be necessary for chronic intracerebroventricular infusion of neuropeptide Y to produce obesity and related abnormalities. Conversely, glucocorticoids inhibited the body weight-lowering effect of leptin. Such dual action of glucocorticoids may occur within the central nervous system, since both neuropeptide Y and leptin act within the hypothalamus. The aim of this study was to determine the effects of glucocorticoids (dexamethasone) given intracerebroventricularly to normal rats on body weight homeostasis and hypothalamic levels of neuropeptide Y and corticotropin-releasing hormone. Continuous central glucocorticoid infusion for 3 days resulted in marked sustained increases in food intake and body weight relative to saline-infused controls. The infusion abolished endogenous corticosterone output and produced hyperinsulinemia, hypertriglyceridemia, and hyperleptinemia, three salient abnormalities of obesity syndromes. Central glucocorticoid infusion also produced a marked decrease in the expression of uncoupling protein (UCP)-1 and UCP-3 in brown adipose tissue and UCP-3 in muscle. Finally, chronic central glucocorticoid administration increased the hypothalamic levels of neuropeptide Y and decreased those of corticotropin-releasing hormone. When the same dose of glucocorticoids was administered peripherally, it resulted in decreases in food intake and body weight, in keeping with the decrease in hypothalamic neuropeptide Y levels. These results suggest that glucocorticoids induce an obesity syndrome in rodents by acting centrally and not peripherally.
Subject(s)
Brain/physiology , Dexamethasone/administration & dosage , Glucocorticoids/administration & dosage , Obesity/chemically induced , Proteins/analysis , Animals , Body Weight/drug effects , Carrier Proteins/metabolism , Corticosterone/antagonists & inhibitors , Corticotropin-Releasing Hormone/metabolism , Eating/drug effects , Homeostasis/drug effects , Hyperinsulinism/chemically induced , Hypertriglyceridemia/chemically induced , Hypothalamus/metabolism , Injections, Intraperitoneal , Ion Channels , Leptin , Membrane Proteins/metabolism , Mitochondrial Proteins , Neuropeptide Y/metabolism , Rats , Rats, Zucker , Uncoupling Protein 1ABSTRACT
Recently findings that marital status is associated with survival in patients with late-stage prostate cancer led to an examination of the generalizability of this association for all cancers. We restricted the investigation to patients with late-stage cancer using population-based data collected from 261,070 patients with late-stage cancer at multiple sites in the United States to determine relations between marital status and survival. After controlling for age, race, and treatment, married patients with cancers of all major primary sites had significantly better survival than single, separated, divorced, or widowed patients. Although single and widowed patients had the poorest prognosis in general, single patients appeared to show the most consistently poor survival across the different types of cancers. Survival differences by marital status were more pronounced in men than in women. This observation raises the possibility that some characteristics associated with being married delay death from cancer. These findings require investigators to ask new questions about the effect of being married and its possible correlates, such as general health status, access to health care, and socioeconomic status. Known correlates of marital status, such as available social support and social isolation also merit attention in relation to these findings.