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1.
Hum Reprod ; 34(1): 127-136, 2019 Jan 01.
Article in English | MEDLINE | ID: mdl-30496407

ABSTRACT

STUDY QUESTION: What are the causal relationships between polycystic ovary syndrome (PCOS) and body mass index (BMI)? SUMMARY ANSWER: Bidirectional Mendelian randomization analyses suggest that increased BMI is causal for PCOS while the reverse is not the case. WHAT IS KNOWN ALREADY: The contribution of obesity to the pathogenesis of PCOS is controversial. To date, published genetic studies addressing this question have generated conflicting results and have not utilized the full extent of known single nucleotide polymorphisms associated with body mass index (BMI). STUDY DESIGN, SIZE, DURATION: This cross-sectional Mendelian randomization (MR) and genetic association study was conducted in 750 individuals of European origin and with PCOS and 1567 BMI-matched controls. PARTICIPANTS/MATERIALS, SETTING, METHODS: Cases and controls were matched for BMI as well as for distribution of weight categories (normal weight, overweight, obese). Two-sample MR using inverse variance weighting (IVW) was conducted using a 92-SNP instrument variable for BMI with PCOS as the outcome, followed by two-sample MR with a 16-SNP instrument variable for PCOS with BMI as the outcome. Sensitivity analyses included MR-Egger and maximum likelihood methods. Secondary analyses assessed associations of genetic risk scores and individual SNPs with PCOS, BMI and quantitative androgen-related and glucose homeostasis-related traits. MAIN RESULTS AND THE ROLE OF CHANCE: Each standard deviation genetically higher BMI was associated with a 4.89 (95% CI 1.46-16.32) higher odds of PCOS. Conversely, genetic risk of PCOS did not influence BMI. Sensitivity analyses yielded directionally consistent results. The genetic risk score of 92 BMI SNPs was associated with the diagnosis of PCOS (OR 1.043, 95% CI 1.009-1.078, P = 0.012). Of the 92 BMI risk variants evaluated, none were associated individually with PCOS after considering multiple testing. The association of FTO SNP rs1421085 with BMI was stronger in women with PCOS (ß = 0.071, P = 0.0006) than in controls (ß = 0.046, P = 0.065). LIMITATIONS, REASONS FOR CAUTION: The current sample size, while providing good power for MR and genetic risk score analyses, had limited power to demonstrate association of individual SNPs with PCOS. Cases and controls were not matched for age; however, this was mitigated by adjusting analyses for age. Dietary and lifestyle data, which could have been used to explore the greater association of the FTO SNP with BMI in women with PCOS, was not available. WIDER IMPLICATIONS OF THE FINDINGS: Increasing BMI appears to be causal for PCOS but having PCOS does not appear to affect BMI. This study used the most comprehensive set of SNPs for BMI currently available. Prior studies using fewer SNPs had yielded conflicting results and may have been confounded because cases and controls were not matched for weight categories. The current results highlight the potential utility of weight management in the prevention and treatment of PCOS. STUDY FUNDING/COMPETING INTEREST(S): National Institutes of Health Grants R01-HD29364 and K24-HD01346 (to R.A.), Grant R01-DK79888 (to M.O.G.), Grant U54-HD034449 (to R.S.L.), Grant U19-HL069757 (to R.M.K.). The funders had no influence on the data collection, analyses or conclusions of the study. No conflict of interests to declare. TRIAL REGISTRATION NUMBER: N/A.


Subject(s)
Alpha-Ketoglutarate-Dependent Dioxygenase FTO/genetics , Body Mass Index , Genetic Predisposition to Disease , Mendelian Randomization Analysis , Polycystic Ovary Syndrome/genetics , Adult , Aged , Case-Control Studies , Cross-Sectional Studies , Female , Genetic Association Studies , Humans , Middle Aged , Polymorphism, Single Nucleotide , Young Adult
2.
J Perinatol ; 30(9): 584-9, 2010 Sep.
Article in English | MEDLINE | ID: mdl-20220763

ABSTRACT

OBJECTIVE: To determine whether 17-alpha hydroxyprogesterone (17-OHPC) alters tumor necrosis factor-alpha (TNF-alpha) production and the expression of cyclooxygenase type 2 (COX-2) in myometrium exposed to lipopolysaccharide (LPS). STUDY DESIGN: Lower segment myometrial biopsies were obtained from non-laboring patients at term. Tissues were cultured in serum-free media with 17-OHPC (1 microM) and LPS (1 microg/ml), either alone or in combination. At 24 h, the production of tumor necrosis factor-alpha (TNF-alpha) and the expression of COX-2 was determined using enzyme linked immunosorbent assay and real-time (RT-PCR). Statistical analysis was performed using non-parametric testing. A P-value of <0.05 was considered significant. RESULT: 17-OHPC had no effect on TNF-alpha production and COX-2 expression when compared with untreated myometrial explants (P=0.61 and P=0.95). LPS induced production of TNF-alpha (P=0.03) and expression of COX-2 (P=0.02). Treatment with 17-OHPC did not block LPS-induced TNF-alpha production (P=0.37) or COX-2 expression (P=0.12). CONCLUSION: In this pilot study, 17-OHPC did not affect the production of TNF-alpha or COX-2 expression in human myometrium.


Subject(s)
Cyclooxygenase 2/metabolism , Hydroxyprogesterones/pharmacology , Myometrium/drug effects , Myometrium/metabolism , Tumor Necrosis Factor-alpha/metabolism , 17 alpha-Hydroxyprogesterone Caproate , Cells, Cultured , Cyclooxygenase 2/genetics , Female , Humans , Lipopolysaccharides , Pregnancy , RNA, Messenger/metabolism
3.
J Neurol Neurosurg Psychiatry ; 71(6): 720-6, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11723190

ABSTRACT

OBJECTIVES: To establish the link between frontal lobe dysfunction and violent and criminal behaviour, based on a review of relevant literature. METHODS: Articles relating evidence of frontal lobe dysfunction with violence or crime were collected through a MEDLINE search using the keyword "frontal lobe" combined with the terms "aggression," "violence," "crime," "antisocial personality disorder," "psychopathy," "impulse control disorders", and "episodic dyscontrol." Reference lists were then searched for additional articles. RESULTS: High rates of neuropsychiatric abnormalities reported in persons with violent and criminal behaviour suggest an association between aggressive dyscontrol and brain injury, especially involving the frontal lobes. The studies reviewed support an association between frontal lobe dysfunction and increased aggressive and antisocial behaviour. Focal orbitofrontal injury is specifically associated with increased aggression. Deficits in frontal executive function may increase the likelihood of future aggression, but no study has reliably demonstrated a characteristic pattern of frontal network dysfunction predictive of violent crime. CONCLUSIONS: Clinically significant focal frontal lobe dysfunction is associated with aggressive dyscontrol, but the increased risk of violence seems less than is widely presumed. Evidence is strongest for an association between focal prefrontal damage and an impulsive subtype of aggressive behaviour.


Subject(s)
Aggression/psychology , Antisocial Personality Disorder/etiology , Brain Diseases/complications , Brain Diseases/psychology , Craniocerebral Trauma/complications , Craniocerebral Trauma/psychology , Criminal Psychology , Frontal Lobe , Neurocognitive Disorders/etiology , Violence/psychology , Antisocial Personality Disorder/diagnosis , Antisocial Personality Disorder/epidemiology , Biological Psychiatry , Craniocerebral Trauma/diagnosis , Criminology , Humans , Magnetic Resonance Imaging , Neurocognitive Disorders/diagnosis , Neurocognitive Disorders/epidemiology , Neuropsychological Tests , Predictive Value of Tests , Research Design/standards , Risk Factors , Tomography, Emission-Computed , Tomography, Emission-Computed, Single-Photon
4.
Epilepsy Behav ; 1(3): 145-9, 2000 Jun.
Article in English | MEDLINE | ID: mdl-12609146
5.
Cancer Chemother Pharmacol ; 41(3): 173-85, 1998.
Article in English | MEDLINE | ID: mdl-9443633

ABSTRACT

The entry of new anticancer treatments into phase I clinical trials is ordinarily based on relatively modest preclinical data. This report defines the battery of preclinical tests important for assessing safety under an Investigational New Drug application (IND) and outlines a basis for extrapolating starting doses of investigational anticancer drugs in phase I clinical trials from animal toxicity studies. Types of preclinical studies for the support of marketing of a new anticancer drug are also discussed. This report addresses differences and similarities in the preclinical development of cytotoxic drugs (including photosensitizers and targeted delivery products), drugs used chronically (chemopreventive drugs, hormonal drugs, immunomodulators), and drugs intended to enhance the efficacy (MDR-reversing agents and radiation/chemotherapy sensitizers) or diminish the toxicity of currently used anticancer therapies. Factors to consider in the design of preclinical studies of combination therapies, alternative therapies, and adjuvant therapies in the treatment of cancer, and to support changes in clinical formulations or route of administration, are also discussed.


Subject(s)
Antineoplastic Agents , Drug Evaluation, Preclinical/methods , Drugs, Investigational , Animals , Antineoplastic Agents/pharmacokinetics , Antineoplastic Agents/therapeutic use , Antineoplastic Agents/toxicity , Clinical Trials, Phase I as Topic , Drug Approval , Drug Evaluation, Preclinical/standards , Drugs, Investigational/pharmacokinetics , Drugs, Investigational/therapeutic use , Drugs, Investigational/toxicity , Humans , United States , United States Food and Drug Administration
6.
Regul Toxicol Pharmacol ; 25(2): 189-93, 1997 Apr.
Article in English | MEDLINE | ID: mdl-9185894

ABSTRACT

The standard approaches for the preclinical development of chronically administered drugs also apply to most respiratory drugs. Modifications from the standard preclinical development plan, however, may be necessary if the drug is administered intranasally or by inhalation. Administration by these routes may result in airway toxicity and the intended patient population is often particularly susceptible. Current and former representatives of the Division of Pulmonary Drug Products (CDER, U.S. FDA) present this article to describe general principles of preclinical development for respiratory drug indications. The article addresses drugs intended for administration by the intranasal or inhalation routes. The article describes the types of studies recommended, considers the initial human dose, and discusses dose-escalation strategies in clinical trials. Other areas of special concern with intranasal or inhalation administration include immunotoxicity, reproductive toxicity, types of dosing apparatus, excipients and extractables, and formulation changes. The approaches described in this article are intended as general information and should be adapted to the scientific considerations and circumstances of a particular drug under development.


Subject(s)
Research , Respiratory System Agents/toxicity , Humans , Research Design
7.
Arch Neurol ; 53(12): 1252-6, 1996 Dec.
Article in English | MEDLINE | ID: mdl-8970456

ABSTRACT

BACKGROUND: We present the signs, symptoms, and radiographic features of 36 children with ischemic infarctions of the basal ganglia, internal capsule, or thalamus. PATIENTS AND METHODS: The series includes 14 males and 22 females ranging in age from newborn to 13 years. Twenty-seven patients were evaluated with computed tomography, 34 with magnetic resonance imaging, 16 with magnetic resonance angiography, and 10 with conventional cerebral angiography. Thirty patients had unilateral lesions (16 left, 14 right) and 6 had bilateral infarctions. RESULTS: The most common presenting symptom was hemiplegia (30 of 36). Other children presented with aphasia (5 of 36), seizures (5 of 36), altered consciousness (5 of 36), and hemisensory changes (5 of 36). Four of 6 patients with bilateral lesions presented with altered mental status, but the location of a unilateral infarction within the thalamus or basal ganglia did not predict the clinical presentation. CONCLUSIONS: The risk factors for basal ganglia infarction in children are diverse, but systemic hypertension does not play a major role in children. The vascular occlusion often occurred in the large arteries, with secondary occlusion of the smaller penetrating arteries. Most children with a single unilateral infarction have a good prognosis.


Subject(s)
Basal Ganglia/blood supply , Basal Ganglia/diagnostic imaging , Cerebral Infarction/diagnostic imaging , Thalamus/blood supply , Thalamus/diagnostic imaging , Adolescent , Cerebral Infarction/complications , Cerebral Infarction/pathology , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Magnetic Resonance Imaging , Male , Radiography
9.
Cancer ; 73(5): 1361-7, 1994 Mar 01.
Article in English | MEDLINE | ID: mdl-8111702

ABSTRACT

BACKGROUND: Small cell carcinoma (SCC) associated with clinical evidence of tumor corticotropin (ACTH) production is common, and management of this syndrome is difficult. The purpose of this retrospective analysis is to describe clinical features, prognosis, and treatment results in patients with SCC and the syndrome of ectopic ACTH secretion to permit formulation of management guidelines for these patients. METHODS: Using tumor registry data and chart review, the authors identified patients with SCC and ectopic ACTH secretion treated over 11 years at two large teaching hospitals. They recorded clinical and laboratory data regarding the patients' tumors and their endocrine syndrome along with results of treatment for the malignancy and the hypercortisolism. RESULTS: Ten patients with SCC and ectopic ACTH secretion were identified. These patients were initially seen with adverse prognostic features, including elevations of serum lactate dehydrogenase and extensive stage disease. Cytotoxic chemotherapy and standard doses of anti-adrenal medications rarely controlled the paraneoplastic syndrome. Bacterial or opportunistic infections, although not neutropenic, developed in most patients. Median survival of patients diagnosed with the paraneoplastic syndrome at the same time as the initial diagnosis of cancer was 4 months. However, three patients whose cortisol secretion was controlled survived longer than 6 months. CONCLUSIONS: Patients with SCC and ectopic ACTH syndrome have a poor prognosis. However, in the minority of patients whose hypercortisolism can be controlled with cytotoxic chemotherapy combined with treatment to inhibit cortisol biosynthesis, effective palliation can be achieved.


Subject(s)
ACTH Syndrome, Ectopic/drug therapy , Carcinoma, Small Cell/drug therapy , Lung Neoplasms/drug therapy , ACTH Syndrome, Ectopic/etiology , Adrenocorticotropic Hormone/blood , Aged , Carcinoma, Small Cell/complications , Carcinoma, Small Cell/mortality , Female , Humans , Hydrocortisone/blood , L-Lactate Dehydrogenase/blood , Lung Neoplasms/complications , Lung Neoplasms/mortality , Male , Middle Aged , Paraneoplastic Syndromes/complications , Prognosis , Retrospective Studies
10.
Cancer ; 71(9): 2813-6, 1993 May 01.
Article in English | MEDLINE | ID: mdl-7682150

ABSTRACT

BACKGROUND: Advanced non-Hodgkin lymphoma (NHL) usually is treated with doxorubicin-based combination chemotherapy. Because doxorubicin is excreted by the biliary route, many authorities recommend alternative initial interventions in patients with NHL causing obstructive jaundice. METHODS: The authors retrospectively reviewed the records of patients at Rochester General Hospital with NHL between 1983 and 1989 with obstructive jaundice at initial diagnosis. RESULTS: Five patients with obstructive jaundice due to intermediate-grade NHL were treated with combination chemotherapy without prior surgical or endoscopic biliary decompression, or radiation therapy. Three received higher doses of doxorubicin than called for in standard dose modification tables. Jaundice was relieved rapidly in all five patients without unexpected toxic effects, and all five patients entered remission (three had partial remission and two complete remission). CONCLUSIONS: Because biliary obstruction resolves rapidly after administration of chemotherapy for this disease, it is possible that standard dose reductions for doxorubicin and vincristine that are appropriate for patients with hepatocellular disease may be excessive for patients with obstructive jaundice resulting from lymphoma. The results of this study indicate that combination chemotherapy is appropriate initial therapy for patients with newly diagnosed NHL with obstructive jaundice.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Cholestasis/drug therapy , Lymphoma, Non-Hodgkin/complications , Adult , Aged , Bleomycin/administration & dosage , Cholestasis/etiology , Cyclophosphamide/administration & dosage , Dexamethasone/administration & dosage , Doxorubicin/administration & dosage , Female , Humans , Leucovorin/administration & dosage , Male , Methotrexate/administration & dosage , Middle Aged , Prednisone/administration & dosage , Retrospective Studies , Vincristine/administration & dosage
13.
Biochemistry ; 29(14): 3536-42, 1990 Apr 10.
Article in English | MEDLINE | ID: mdl-2354150

ABSTRACT

Human neutrophil cathepsin G or bovine chymotrypsin proteolytically cleaved human alpha-thrombin at the B-chain Trp148-Thr149 bond generating a new form, zeta-thrombin. While incubation of alpha-thrombin with cathepsin G at pH 7.4 and 37 degrees C resulted in a partial loss of fibrinogen clotting activity, 86 +/- 13% of the clotting activity and 99 +/- 16% of the active sites titratable with p-nitrophenyl p-guanidinobenzoate were retained upon controlled passage of alpha-thrombin through chymotrypsin-Sepharose 4B at pH 6.2 or 7.4 and 24 degrees C (n = 15). Kinetic parameters for H-D-hexahydrotyrosyl-Ala-Arg p-nitroanilide were Km = 1.52 +/- 0.60 vs 1.32 +/- 0.18 microM and kcat = 51.9 +/- 2.9 vs 35.8 +/- 6.4 s-1 with alpha-thrombin vs chymotrypsin-prepared zeta-thrombin (n = 4 vs 3), respectively (I = 0.15 M, pH 7.4, and 24 degrees C). Some 95% of the clotting activity was lost when zeta-thrombin was passed through trypsin-Sepharose 4B under conditions for converting alpha- to nonclotting beta- and subsequently gamma-thrombin. The resulting gamma-like thrombins eluted bimodally with 260 and 310 mM NaCl when applied to Amberlite CG-50 resin [cross-linked poly(methylacrylic acid)] developed with a linear salt gradient in 50 mM Tris at pH 7.4 and 24 degrees C. These elution peaks correspond to 240, 330, and 350 mM NaCl for gamma-, alpha-, and zeta-thrombin, respectfully, implying that the anion-binding exosite is partially destroyed in gamma-like thrombins but is intact in zeta-thrombin.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Blood Coagulation , Cathepsins/blood , Chymotrypsin/metabolism , Fibrinogen/metabolism , Neutrophils/enzymology , Thrombin/metabolism , Amino Acid Sequence , Animals , Cathepsin G , Cattle , Humans , Hydrolysis , Kinetics , Macromolecular Substances , Molecular Sequence Data , Oligopeptides/chemical synthesis , Oligopeptides/pharmacology , Protease Inhibitors/chemical synthesis , Protease Inhibitors/pharmacology , Sequence Homology, Nucleic Acid , Serine Endopeptidases , Substrate Specificity
15.
Blood ; 74(5): 1627-34, 1989 Oct.
Article in English | MEDLINE | ID: mdl-2477083

ABSTRACT

Polymorphonuclear leukocytes (PMN) when activated release products that can potentially injure endothelial cells or alter endothelial function. Exposure of cultured human umbilical vein endothelial cells to cathepsin G and elastase isolated from human PMN at concentrations reached in vivo (100 ng/mL to 10 micrograms/mL) selectively inhibited thrombin-induced prostacyclin production and the thrombin-induced rise in cytosolic free calcium ([Ca++]i) concentration. These proteases also blocked thrombin-induced release of arachidonic acid from prelabeled endothelial cells (EC). In contrast, induction of prostacyclin (PGI2) production by arachidonate, histamine, or the calcium ionophore A23187 was not altered by treatment of EC with these proteases. The effects of the proteases were concentration-dependent, were blocked by serum or serum protease inhibitors, and were reversed when the endothelial cells were further cultured for 24 hours in the absence of the proteases. Elastase, but not cathepsin G, also produced detachment of endothelial cells. Thus, the major leukocyte proteases selectively suppress thrombin-induced prostacyclin production by human vascular endothelial cells and may alter the hemostatic balance at sites of PMN activation.


Subject(s)
Cathepsins/blood , Endothelium, Vascular/metabolism , Epoprostenol/biosynthesis , Leukocytes/enzymology , Pancreatic Elastase/blood , Thrombin/physiology , Arachidonic Acid , Arachidonic Acids/metabolism , Calcimycin/pharmacology , Calcium/metabolism , Cathepsin G , Cathepsins/isolation & purification , Cells, Cultured , Endothelium, Vascular/drug effects , Humans , Kinetics , Neutrophils/enzymology , Pancreatic Elastase/isolation & purification , Serine Endopeptidases , alpha 1-Antitrypsin/physiology , alpha-Macroglobulins/physiology
17.
Blood ; 69(3): 813-9, 1987 Mar.
Article in English | MEDLINE | ID: mdl-3101765

ABSTRACT

During blood coagulation, polymorphonuclear leukocytes release elastase in amounts that can exceed 100 nmol/L. We therefore studied the interaction between human leukocyte elastase and human alpha-thrombin. Elastase cleaved the thrombin B chain (Ala 150-Asn 151) near the gamma-cleavage site, resulting in two fragments held together by noncovalent interactions. The NH2-terminal fragment (FI), mol wt approximately 18,000, was disulfide-linked to the thrombin A chain. The COOH-terminal fragment (FII), mol wt approximately 13,000, contained the active-site serine and formed a covalent bond with antithrombin III. Heparin accelerated proteolysis of alpha-thrombin by elastase. Proteolyzed alpha-thrombin (T theta) retained full amidolytic activity; however, the concentration of T theta causing 50% maximal platelet aggregation and adenosine triphosphate (ATP) release was 7.9 nmol/L (1.1 nmol/L for alpha-thrombin and 220 nmol/L for gamma-thrombin). Fibrinogen clotting activity of T theta and gamma-thrombin was 32% and 1% that of alpha-thrombin, respectively. Elastase released during the coagulation process may modulate thrombin activity. In addition, elastase-modified thrombin may be a useful probe of the structure and function of the gamma-cleavage region.


Subject(s)
Blood Coagulation/drug effects , Fibrinogen/metabolism , Neutrophils/enzymology , Pancreatic Elastase/metabolism , Platelet Aggregation/drug effects , Thrombin/metabolism , Amino Acid Sequence , Antithrombin III/metabolism , Humans , Serine/metabolism , Thrombin/pharmacology
18.
Am J Clin Oncol ; 10(1): 47-9, 1987 Feb.
Article in English | MEDLINE | ID: mdl-3825992

ABSTRACT

Twenty-two patients with metastatic colorectal cancer were treated with a regimen of 5-fluorouracil 600 mg/m2 (maximum, 1.0 g) i.v./week and folic acid 140 mg/m2 i.v. given 1 h prior to the 5-FU. This study was undertaken in an attempt to confirm the in vitro finding that inhibition of thymidylate synthetase by 5-fluorouracil is prolonged by the presence of folates. There were four partial responses (18%) with mean duration 4 months. Dose-limiting toxicity was enteritis, seen in 12 patients (58%), and causing hospitalization in seven patients. Enteritis was shown to be due to the folic acid in most patients. Two patients died from leukopenia, enteritis, and sepsis. Mean serum folate levels at the time of 5-FU injection were 36 microM. This regimen is no more effective than 5-FU alone and has significantly more serious toxicity. Further investigation of this regimen is not recommended.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Colonic Neoplasms/drug therapy , Fluorouracil/administration & dosage , Folic Acid/administration & dosage , Antineoplastic Combined Chemotherapy Protocols/adverse effects , Diarrhea/chemically induced , Female , Folic Acid/blood , Humans , Male , Middle Aged , Neoplasm Metastasis
19.
Cancer Res ; 46(2): 798-806, 1986 Feb.
Article in English | MEDLINE | ID: mdl-3940644

ABSTRACT

We tested the ability of serum-free media to support the in vitro growth of human non-small cell lung carcinoma. A medium containing insulin, transferrin, sodium selenite, hydrocortisone, epidermal growth factor, and bovine serum albumin (1 mg/ml) with serum precoating of culture dishes (modified LA medium) supported three previously established cell lines of non-small cell lung cancer and prevented fibroblast proliferation in fresh tumor specimens but did not support long term tumor cell growth from fresh specimens. We added triiodothyronine, sodium pyruvate, and additional glutamine, insulin, and epidermal growth factor to modified LA medium, precoated with fibronectin and collagen instead of serum, and deleted bovine serum albumin, defining a new medium called ACL-3. ACL-3 medium alone supported the short term growth of 10 of 12 cell lines and the soft agarose cloning of 9 of 12 cell lines tested, and ACL-3 supplemented by an optimal concentration of bovine serum albumin (5 mg/ml) supported the long term growth of 10 of 12 cell lines tested. Moreover, we have grown tumor cells for more than 6 months from 11 of 33 (33%) consecutive fresh clinical specimens of human lung adenocarcinoma in ACL-3 with bovine serum albumin. ACL-3 medium provides a defined environment for the study of growth factor requirements of human non-small cell lung cancer and enhances our ability to grow human lung cancer, particularly adenocarcinoma, in vitro.


Subject(s)
Adenocarcinoma/pathology , Culture Media , Lung Neoplasms/pathology , Carcinoma, Squamous Cell/pathology , Cell Division/drug effects , Cell Line , Culture Techniques , Extracellular Matrix/physiology , Growth Substances/pharmacology , Humans , Sepharose
20.
Anal Biochem ; 147(2): 382-6, 1985 Jun.
Article in English | MEDLINE | ID: mdl-4014681

ABSTRACT

A technique for the detection of nanogram amounts of protein blotted onto nitrocellulose membranes has been developed using nonradioactive probes. Protein transferred to nitrocellulose membranes is detected by a specific antibody followed by incubation with biotinylated anti-antibody. After addition of streptavidin-acid phosphatase complex, incubation with fast violet B salt produces sharp magenta bands. This method allows detection of bands containing less than 20 ng of protein. The procedure does not use radioactive or carcinogenic materials.


Subject(s)
Acid Phosphatase , Avidin , Collodion , Ovalbumin/analogs & derivatives , Proteins/analysis , Animals , Biotin , Color , Horseradish Peroxidase , Humans , Immunoassay , Rabbits
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