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2.
Mol Genet Metab ; 71(4): 591-8, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11136551

ABSTRACT

We report three novel activating mutations in the calcium-sensing receptor (CASR) that are responsible for autosomal dominant hypocalcemia (ADH) in three unrelated families. Each mutation involves a missense substitution resulting in a nonconservative amino acid alteration, P221L, E228Q, and Q245R. These mutations were observed in affected family members, but not in unaffected family members or in unrelated control samples. All three mutations are clustered in the extracellular domain of the CASR in a region dominated by negatively charged amino acids. Each mutant and wild-type receptor was expressed in Cos-1 cells. A luciferase reporter gene assay was utilized to detect the level of receptor activity by utilizing a protein kinase C-activated promoter to drive the production of luciferin, the reporter gene product. All three mutant receptors exhibited an increased sensitivity to calcium at all concentrations tested when compared to the wild-type receptor, supporting the hypothesis that these are activating mutations and are responsible for the ADH phenotype in these families. The data presented in this study suggest the importance of this highly negatively charged region of the extracellular domain in normal CASR function.


Subject(s)
Genes, Dominant/genetics , Hypocalcemia/genetics , Mutation/genetics , Receptors, Cell Surface/genetics , Adult , Animals , Base Sequence , COS Cells , Calcium/metabolism , Child , DNA Mutational Analysis , Female , Genes, Reporter , Humans , Hypocalcemia/enzymology , Hypocalcemia/metabolism , Infant , Male , Molecular Sequence Data , Pedigree , Polymorphism, Restriction Fragment Length , Polymorphism, Single-Stranded Conformational , Protein Kinase C/metabolism , Receptors, Calcium-Sensing , Receptors, Cell Surface/metabolism , Transfection
3.
Mol Cell Biol ; 14(9): 5961-74, 1994 Sep.
Article in English | MEDLINE | ID: mdl-8065329

ABSTRACT

The bone morphogenetic proteins (BMPs) are a group of transforming growth factor beta (TGF-beta)-related factors whose only receptor identified to date is the product of the daf-4 gene from Caenorhabditis elegans. Mouse embryonic NIH 3T3 fibroblasts display high-affinity 125I-BMP-4 binding sites. Binding assays are not possible with the isoform 125I-BMP-2 unless the positively charged N-terminal sequence is removed to create a modified BMP-2, 125I-DR-BMP-2. Cross-competition experiments reveal that BMP-2 and BMP-4 interact with the same binding sites. Affinity cross-linking assays show that both BMPs interact with cell surface proteins corresponding in size to the type I (57- to 62-kDa) and type II (75- to 82-kDa) receptor components for TGF-beta and activin. Using a PCR approach, we have cloned a cDNA from NIH 3T3 cells which encodes a novel member of the transmembrane serine/threonine kinase family most closely resembling the cloned type I receptors for TGF-beta and activin. Transient expression of this receptor in COS-7 cells leads to an increase in specific 125I-BMP-4 binding and the appearance of a major affinity-labeled product of approximately 64 kDa that can be labeled by either tracer. This receptor has been named BRK-1 in recognition of its ability to bind BMP-2 and BMP-4 and its receptor kinase structure. Although BRK-1 does not require cotransfection of a type II receptor in order to bind ligand in COS cells, complex formation between BRK-1 and the BMP type II receptor DAF-4 can be demonstrated when the two receptors are coexpressed, affinity labeled, and immunoprecipitated with antibodies to either receptor subunit. We conclude that BRK-1 is a putative BMP type I receptor capable of interacting with a known type II receptor for BMPs.


Subject(s)
Caenorhabditis elegans Proteins , Protein Serine-Threonine Kinases/genetics , Proteins/metabolism , Receptors, Cell Surface/genetics , Receptors, Growth Factor/genetics , Receptors, Transforming Growth Factor beta , 3T3 Cells , Amino Acid Sequence , Animals , Base Sequence , Bone Morphogenetic Protein Receptors, Type I , Bone Morphogenetic Proteins , Cloning, Molecular , Gene Expression , Helminth Proteins/metabolism , Mice , Molecular Sequence Data , Oligonucleotide Probes/chemistry , RNA, Messenger/genetics , Receptor Protein-Tyrosine Kinases/metabolism , Recombinant Proteins , Sequence Alignment , Sequence Homology, Amino Acid , Tissue Distribution
4.
Spine (Phila Pa 1976) ; 18(16): 2523-4, 1993 Dec.
Article in English | MEDLINE | ID: mdl-8303458

ABSTRACT

Herpes zoster-related radiculopathy usually can be easily diagnosed in the presence of cutaneous lesions. Before development of the skin rash, the diagnosis may be in doubt, particularly if motor symptoms and signs are a major clinical feature. We report a patient with herpes zoster-related radiculopathy whose clinical features mimicked other spinal disorders.


Subject(s)
Herpes Zoster/diagnosis , Paralysis/etiology , Peripheral Nervous System Diseases/microbiology , Aged , Diagnosis, Differential , Female , Herpes Zoster/complications , Humans , Leg/innervation , Peripheral Nervous System Diseases/complications , Time Factors
5.
Med Sci Sports Exerc ; 25(10): 1185-91, 1993 Oct.
Article in English | MEDLINE | ID: mdl-8231765

ABSTRACT

The purpose of this study was to evaluate the validity and reliability of the Futrex 5000A near infrared (NIR) spectrophotometer for the assessment of body composition in children and adolescents. Forty-eight subjects (24 boys and 24 girls) with a mean age of 12.7 +/- 2.7 yr underwent three methods of body composition testing: NIR, densitometry by hydrostatic weighing (HYDRO), and skinfold anthropometry (SKF). Percent body fat (%BF) and fat free mass (FFM) derived from the Lohman's age-adjusted Siri equation served as the criterion. Within session test-retest reliability was determined for the NIR device for all subjects and between-week reliability was evaluated for all test methods in 14 subjects. Based on the excellent within- and between-session reliability (ICC ranged from 0.907-0.999), the system offers the potential of obtaining longitudinal data in growth and development studies. However, the significant mean differences, moderate correlations with the criterion (r = 0.62-0.71) and inflated standard errors of estimate (SEE = 4.9-5.5% BF, 2.2-2.9 kg FFM) and total prediction errors (TE = 5.5-8.0% BF, 2.7-3.7 kg FFM) indicate that refinement of prediction equations is needed to establish the measurement validity. Continued research with expanded populations is needed to further demonstrate and evaluate the utility of this device.


Subject(s)
Body Composition , Spectrophotometry, Infrared/methods , Adipose Tissue , Adolescent , Adult , Age Factors , Analysis of Variance , Body Mass Index , Body Weight , Child , Densitometry/methods , Female , Humans , Male , Reproducibility of Results , Sex Factors , Skinfold Thickness
6.
Med Sci Sports Exerc ; 25(9): 1070-6, 1993 Sep.
Article in English | MEDLINE | ID: mdl-8231777

ABSTRACT

Prior to sexual maturation, children and adolescents have more water and less bone mineral content than adults, resulting in less dense fat-free body mass (FFM). This suggests that previously established adult skinfold/density equations are inappropriate for use with children and adolescents for the prediction of body fatness (%BF) and FFM. To overcome this problem, Slaughter and colleagues have introduced new skinfold (SKF) equations that take into account the changing density of FFM in children and adolescents as they mature. The purpose of our study was to cross-validate a select set of the Slaughter SKF equations by comparing them with a criterion measurement (Lohman's Siri age-adjusted body density equation) in 122 subjects ranging in sexual maturation from pre- to post-pubescent and ranging in age from 8-17. Our cross-validation found very high intraclass (reliability) correlations (ICCs = 0.98-0.99) and high validity correlations (rs = 0.79-0.99). The standard errors of the estimate for %BF ranged from 3.5-4.6% and total errors for %BF ranged from 3.6-4.6%. The Slaughter equation using tricep and calf SKF for females was significantly different (P < 0.05) from the criterion measure in its prediction of %BF. In males, there was an interaction between the SKF equation and subject maturation level. The data indicate that the Slaughter SKF equations hold promise for estimating body composition in children and adolescents but are still in need of refinement.


Subject(s)
Skinfold Thickness , Adipose Tissue , Adolescent , Body Composition , Body Constitution , Body Mass Index , Body Weight , Child , Female , Humans , Male , Muscles/anatomy & histology , Observer Variation , Reproducibility of Results , Residual Volume , Sex Factors
7.
J Clin Endocrinol Metab ; 77(3): 725-30, 1993 Sep.
Article in English | MEDLINE | ID: mdl-7690363

ABSTRACT

During puberty, plasma insulin levels increase, and insulin sensitivity decreases along with multiple other physical and hormonal changes. To determine 1) the time course of the decrease in insulin sensitivity in relationship to Tanner stage of genital development, and 2) how this change relates to changes in GH secretion, insulin-like growth factor-I (IGF-I), IGF-binding protein-3, and gonadal steroid secretion, we studied 58 healthy children and adolescents (34 males and 24 females; age 7-15 yr) using overnight GH sampling and frequently sampled iv glucose tolerance tests. The insulin sensitivity index (ISI) was calculated using the program MINMOD. ISI differed significantly by Tanner stage (P < 0.05, by analysis of variance) with a decrease from Tanner stage 1 to 2 (P < 0.05). IGF-I and IGF-binding protein-3 followed opposite patterns to ISI, with lower levels in Tanner stage 1 than in stages 2-5 (P < 0.05). Mean GH levels did not increase until Tanner stage 4 (P < 0.05) and then fell during Tanner stage 5. Multiple linear regression analysis revealed negative relationships among ISI, IGF-I, and body mass index. No relationship was found with GH. We conclude that the pubertal change in ISI is not necessarily associated with increased GH secretion, but is associated with increased GH peripheral effect, as indicated by the relationship between ISI and IGF-I.


Subject(s)
Insulin/pharmacology , Puberty/physiology , Adolescent , Body Mass Index , Carrier Proteins/metabolism , Child , Estradiol/blood , Female , Glucose , Growth Hormone/metabolism , Humans , Insulin/blood , Insulin-Like Growth Factor Binding Proteins , Insulin-Like Growth Factor I/metabolism , Male , Regression Analysis , Sex Characteristics , Testosterone/blood
8.
Phys Ther ; 73(5): 320-8, 1993 May.
Article in English | MEDLINE | ID: mdl-8469716

ABSTRACT

BACKGROUND AND PURPOSE: The reliability and validity of measurements obtained with two bioelectrical impedance analyzers (BIAs), an RJL Systems model BIA-103 and a Berkeley Medical Research BMR-2000, were investigated using the manufacturers' prediction equations for the assessment of fat-free mass (FFM) (in kilograms) in children and adolescents. SUBJECTS: Forty-seven healthy children and adolescents (23 male, 24 female), ranging in age from 8 to 20 years (mean = 12.1, SD = 2.3), participated. METHODS: In the context of a repeated-measures design, the data were analyzed according to gender and maturation (Tanner staging). Hydrostatic weighing (HYDRO) and Lohman's Siri age-adjusted body density prediction equation served as the criteria for validating the BIA-obtained measurements. RESULTS: High intraclass correlation coefficients (ICC > or = .987) demonstrated good test-retest (between-week) measurement reliability for HYDRO and both BIA methods. Between-method (HYDRO versus BIA) correlation coefficients were high for both boys and girls (r > or = .97). The standard errors of estimate (SEEs) for FFM were slightly larger for boys than for girls and were consistently smaller for the RJL system than for the BMR system (RJL SEE = 1.8 kg for boys, 1.3 kg for girls; BMR SEE = 2.4 kg for boys, 1.9 kg for girls). The coefficients of determination were high for both BIA methods (r2 > or = .929). Total prediction errors (TEs) for FFM showed similar between-method trends (RJL TE = 2.1 kg for boys, 1.5 kg for girls; BMR TE = 4.4 kg for boys, 1.9 kg for girls). DISCUSSION AND CONCLUSION: This study demonstrated that the RJL BIA with the manufacturer's prediction equations can be used to reliably and accurately assess FFM in 8- to 20-year-old children and adolescents. The prediction of FFM by the BMR system was acceptable for girls, but significant overprediction of FFM for boys was noted.


Subject(s)
Body Composition , Electric Impedance , Adolescent , Adult , Child , Female , Humans , Male , Reproducibility of Results
9.
Arch Dermatol ; 128(10): 1345-8, 1992 Oct.
Article in English | MEDLINE | ID: mdl-1417021

ABSTRACT

BACKGROUND AND DESIGN: Increases in sebaceous gland activity are often the earliest sign of the approach of puberty in children. These increases have been attributed to increases in the secretion of adrenal androgens, but the supporting data are sparse and are based on measurements of urinary, rather than serum, androgen concentrations. In this study, we examined sebum composition, serum levels of dehydroepiandrosterone sulfate, and pubertal stage in 111 boys and girls, aged 2 to 15 years. Sebum composition was evaluated by measuring the ratio of wax esters/(cholesterol + cholesterol esters), a ratio known to increase with increasing sebaceous gland activity. RESULTS: Both wax esters/(cholesterol + cholesterol esters) ratios and serum dehydroepiandrosterone sulfate levels began to increase in children 7 to 10 years old. These changes occurred in many children before the appearance of any physical signs of puberty. Wax esters/(cholesterol + cholesterol esters) ratios were correlated with dehydroepiandrosterone sulfate levels in both boys and girls. In prepubertal children, the regression lines passed through the origin. In subjects who were in early or late puberty, the y intercepts of the regression lines had positive values. CONCLUSION: Adrenal androgens appear to be the major determinants of sebaceous gland activity during the prepubertal period and to be additive to another hormone or hormones during puberty.


Subject(s)
Dehydroepiandrosterone/analogs & derivatives , Sebaceous Glands/metabolism , Adolescent , Child , Child, Preschool , Cholesterol/analysis , Cholesterol Esters/analysis , Cohort Studies , Dehydroepiandrosterone/blood , Dehydroepiandrosterone Sulfate , Female , Humans , Male , Sebum/chemistry , Sebum/metabolism , Waxes/analysis
11.
Pediatr Neurol ; 8(3): 229-31, 1992.
Article in English | MEDLINE | ID: mdl-1622523

ABSTRACT

Valproic acid has previously been demonstrated to decrease the secretion of multiple pituitary hormones; however, clinical symptoms associated with decreased hormone secretion have not been described. A girl is reported with complex partial seizures who was treated with valproic acid from age 10 years, 7 months to 12 years, 1 month and during this time had an arrest of both growth and secondary sexual development. Two months after discontinuing valproic acid she had resumption of both pubertal growth and maturation. A prospective controlled study is needed to determine more precisely the effects of valproic acid on growth and development.


Subject(s)
Epilepsy, Complex Partial/drug therapy , Puberty/drug effects , Valproic Acid/adverse effects , Body Height/drug effects , Body Weight/drug effects , Child , Evoked Potentials/drug effects , Female , Humans , Psychosexual Development/drug effects , Valproic Acid/administration & dosage
12.
J Clin Endocrinol Metab ; 74(5): 1206-9, 1992 May.
Article in English | MEDLINE | ID: mdl-1569169

ABSTRACT

The development of GnRH analogs (GnRHa) has made it possible to treat children with central precocious puberty (CPP). This treatment may prevent adult short stature due to premature epiphyseal fusion. Achievement of this goal, however, is dependent upon adequate suppression of gonadal steroid production as a result of GnRHa-induced pituitary desensitization and decreased gonadotropin release. A depot formulation of a GnRHa [leuprolide acetate (dLA)] is being used by many clinicians for the treatment of CPP, but studies to establish the optimal dose of dLA have not been performed. In this study we evaluated the effectiveness of dLA (7.5 mg, im, every 4 weeks). Six children (7-10 yr old) with CPP treated with dLA were assessed clinically and divided into two groups: A (incompletely suppressed) and B (well suppressed). Each group had overnight blood sampling and a GnRH stimulation test the following morning. LH pulses were analyzed and compared to 11 normal prepubertal children. Mean LH concentration, LH curve area, LH pulse frequency, and mean LH pulse amplitude were significantly greater (P less than 0.03) in group A than in group B or the normal prepubertal children. There was no significant difference among the three groups in GnRH-stimulated peak LH release. These results indicate that dLA (7.5 mg, im, every 4 weeks) does not produce complete desensitization in all children with CPP and suggest that overnight monitoring of LH release is more sensitive than GnRH stimulation testing for the assessment of dLA dose adequacy.


Subject(s)
Leuprolide/administration & dosage , Puberty, Precocious/drug therapy , Child , Delayed-Action Preparations , Dose-Response Relationship, Drug , Female , Gonadotropin-Releasing Hormone/pharmacology , Humans , Luteinizing Hormone/metabolism , Male , Puberty, Precocious/metabolism , Time Factors
13.
FEBS Lett ; 300(2): 149-52, 1992 Mar 30.
Article in English | MEDLINE | ID: mdl-1373390

ABSTRACT

We have isolated a portion of the canine gene encoding the orphan receptor RDC1 [1]. The complete coding sequence is contained in a single exon, and an intron divides the 5' untranslated region of RDC1 mRNA. The RDC1 protein is 94% homologous to the gene product of GPRN1, which has been proposed to serve as a VIP receptor when expressed in CHO-K1 and COS-7 cells (Sreedharan, S.P. et al. (1991) Proc. Natl. Acad. Sci. USA 88, 4986-4990). Northern analysis indicates that CHO-K1 cells endogenously express a 2.1 kb RDC1 mRNA. However, while CHO-K1 cells possess detectable low affinity [125I]VIP binding sites, VIP binding is not altered in membranes of CHO-K1 cells expressing varying amounts of the RDC1 gene construct. Further, endogenous VIP binding is not increased by transient expression of RDC1 in COS-7 cells. Taken together, the data suggest that RDC1 is not a canine homolog of the proposed VIP receptor.


Subject(s)
Receptors, Gastrointestinal Hormone/genetics , Vasoactive Intestinal Peptide , Animals , Base Sequence , Blotting, Northern , CHO Cells , Cell Line , Cloning, Molecular , Cricetinae , Dogs , GTP-Binding Proteins/metabolism , Humans , Molecular Sequence Data , RNA/isolation & purification , Receptors, Gastrointestinal Hormone/metabolism , Receptors, Vasoactive Intestinal Peptide , Sequence Homology, Nucleic Acid , Transfection
14.
J Biol Chem ; 267(4): 2274-81, 1992 Feb 05.
Article in English | MEDLINE | ID: mdl-1733935

ABSTRACT

In a preliminary study we demonstrated that the formation of the epidermal growth factor (EGF) receptor-ligand complex requires the participation of the highly conserved arginine 41 side chain of the growth factor peptide (Engler, D.A., Montelione, G.T., and Niyogi, S.K. (1990) FEBS Lett. 271, 47-50). In an attempt to gain further insight into the nature of this interaction(s), we used both site-directed mutagenesis and chemical modification reagents to produce human EGF (hEGF) analogues with altered chemical properties of the residue 41 side chain. Eight mutant analogues of hEGF were generated, substituting arginine 41 with lysine, glutamine, isoleucine, tyrosine, glycine, alanine, aspartate, or glutamate. Although each of the mutant analogues was able to displace wild-type hEGF fully in receptor competition binding assays, affinity of the receptor for the mutants was substantially reduced, varying from 0.4 to less than 0.01% of that observed for wild-type growth factor. At sufficiently high concentrations these mutants were able to stimulate DNA synthesis in mouse keratinocytes. Substitution of lysine for arginine 41 reduced the receptor affinity 250-fold from that observed for wild type, despite retention of the positive electrostatic charge. The lysine substitution leaves a reactive amine at position 41 and made it possible, using amine-specific chemical modification reagents, to produce selected arginine homologues that were tested for their effects on receptor binding, receptor tyrosine kinase activation, and stimulation of DNA synthesis in mouse keratinocytes. The reaction of lysine 41 with methyl acetimidate resulted in a lysineacetamidine product which only partially restored activity of the lysine hEGF mutant. However, reaction with O-methylisourea resulted in generation of an arginine 41 homologue (homoarginine) which restored full activity. The results indicate that the chemical properties inherent in the guanidinium group of the arginine 41 side chain of hEGF are responsible for optimal receptor-ligand association.


Subject(s)
Arginine/metabolism , Epidermal Growth Factor/metabolism , Guanidines/metabolism , Amino Acids/genetics , Animals , Binding, Competitive , Epidermal Growth Factor/drug effects , Epidermal Growth Factor/genetics , ErbB Receptors/metabolism , Guanidine , Humans , Keratinocytes/cytology , Keratinocytes/metabolism , Mice , Mice, Inbred BALB C , Mutagenesis, Site-Directed
15.
Pediatr Radiol ; 22(7): 481-4, 1992.
Article in English | MEDLINE | ID: mdl-1491902

ABSTRACT

Cranial magnetic resonance imaging was performed in 17 children with central precocious puberty (CPP) and 19 aged-matched controls to compare the appearance of the pituitary gland. Gland size was measured on T1-weighted sagittal and coronal images. The gland was graded according to the concavity or convexity of the upper surface, and the signal intensity of the gland was assessed visually. The mean pituitary volume in 13 CPP children without hypothalamic tumor (292.6 mm3) was significantly greater than that in normal controls (181.35 mm3). The mean volume for the four CPP children with hypothalamic tumor was smaller (145.0 mm3). Compared to controls, the upper pituitary surface in CPP patients appeared convex in a higher proportion. The anterior pituitary was isointense to pons in all patients and controls. Although the posterior pituitary bright spot was present in 14 controls and 11 CPP patients, none with hypothalamic tumor showed it.


Subject(s)
Magnetic Resonance Imaging , Puberty, Precocious/diagnosis , Astrocytoma/complications , Astrocytoma/diagnosis , Child , Child, Preschool , Female , Hamartoma/complications , Hamartoma/diagnosis , Humans , Hypothalamic Neoplasms/complications , Hypothalamic Neoplasms/diagnosis , Male , Pituitary Gland/pathology , Puberty, Precocious/complications
16.
Biol Bull ; 182(2): 165-166, 1992 Apr.
Article in English | MEDLINE | ID: mdl-29303669
17.
Mol Cell Biol ; 11(5): 2425-31, 1991 May.
Article in English | MEDLINE | ID: mdl-1850095

ABSTRACT

The third disulfide loop (amino acids 33 to 42) of human epidermal growth factor (hEGF) encompasses the region of highest amino acid conservation among all of the EGF-like family of molecules. The importance of some of these highly conserved residues for the maintenance of biological activity, especially the aromatic amino acid tyrosine at position 37, has until now been considered essential on the basis of previous studies with the EGF-like molecule transforming growth factor alpha. Variants at the Tyr-37 position of hEGF were constructed by site-directed mutagenesis. The substituting amino acids were phenylalanine, histidine, serine, alanine, aspartic acid, arginine, and glycine. The variants were tested for their ability to competitively displace native [125I]hEGF from its receptor and to stimulate the protein-tyrosine kinase activity of the receptor; the order of efficacy of substituting amino acids was Phe greater than His greater than Ser greater than Ala greater than Asp greater than Arg greater than Gly in both assays. All were effective, with no or only moderate reduction in potency, in stimulating the incorporation of [3H]thymidine into acid-insoluble material of quiescent mouse A31 cells. Only Tyr-37----Ala, Tyr-37----Arg and Tyr-37----Gly were slightly less potent in the cell assay. Thus, neither tyrosine nor another aromatic amino acid at position 37 in hEGF is essential for full biological activity.


Subject(s)
Epidermal Growth Factor/genetics , Amino Acid Sequence , Animals , Base Sequence , Binding, Competitive , Epidermal Growth Factor/metabolism , Epidermal Growth Factor/pharmacology , ErbB Receptors/metabolism , Humans , Kinetics , Molecular Sequence Data , Mutagenesis, Site-Directed , Oligonucleotide Probes/chemical synthesis , Protein-Tyrosine Kinases/drug effects , Protein-Tyrosine Kinases/metabolism , Restriction Mapping , Sequence Homology, Nucleic Acid
18.
Health Phys ; 60(5): 657-60, 1991 May.
Article in English | MEDLINE | ID: mdl-2019496

ABSTRACT

Fluorodeoxyglucose labeled with 18F (18F-FDG) is the most commonly used radiopharmaceutical in positron emission tomography (PET). Fluorine-18-labeled FDG is used as a diagnostic tool in PET studies to monitor the physiology of the brain, diagnose heart function and disease, and to image cancerous tumors. At the University of California, Los Angeles (UCLA), three cyclotrons produce [18F]-fluoride ion using 18O-enriched water targets. Fluorine-18, which has a half-life of 109.8 min, is produced using an 18O(p.n.)18F reaction and is chemically processed to yield 18F-FDG. This study presents data which demonstrate that during the radiochemical processes involved in the production of 18F-FDG, gaseous effluent containing 18F is released. Forty cyclotron production runs with average end of cyclotron bombardment activities of 15.9 +/- 1.88 GBq (430 +/- 50.8 mCi) and end of radiochemical synthesis activities of 5.40 +/- 1.27 GBq (146 +/- 34.3 mCi) yield 18F gaseous effluent releases ranging from 0 to 2560 MBq (0 to 69.2 mCi) with a mean of 437 MBq (11.8 mCi). Temporal correlation of the 18F gaseous releases during 18F-FDG radiochemical production has tied the 18F release to the addition of the glucose precursor (mannotriflate) and ethyl ether in the radiochemical processing. The results are presented in terms of activities released and dilution factors required from the release stack point to maintain controlled (occupational) and uncontrolled (public) area limits in accordance with the recommendations of the International Commission on Radiological Protection and the regulatory requirements of the federal government.


Subject(s)
Air Pollutants, Radioactive/analysis , Deoxyglucose/analogs & derivatives , Fluorine Radioisotopes/analysis , Environmental Exposure , Fluorodeoxyglucose F18 , Isotope Labeling , Occupational Exposure , Particle Accelerators
19.
Am J Physiol ; 259(1 Pt 1): C84-91, 1990 Jul.
Article in English | MEDLINE | ID: mdl-2164784

ABSTRACT

Putrescine is taken up by confluent pig kidney (LLC-PK1) cells at roughly equal rates over both Na(+)-dependent and Na(+)-independent pathways. The former is sensitive to 1 mM amiloride, but the latter is not. Uptake rates are similar at both the apical and basolateral surfaces. The principal fate of the putrescine is oxidative deamination, yielding a product that appears to be either gamma-aminobutyraldehyde or delta 1-pyrroline. Most of the remainder is converted to products tentatively identified as spermidine, spermine, or another unidentified product; these products as well as putrescine itself are lost from the cell at either surface. Changing the extracellular pH in the range of 6.8-8.0 has no affect on putrescine uptake. Cells acidified to intracellular pH 6.8 show a reduced capacity to incorporate radioactivity, an effect that may be due to inhibition of diamine oxidase. Depletion of ATP stores by treating cells with 2-deoxy-D-glucose and NaN3 does not reduce putrescine uptake, suggesting that the mechanism is not a primary active transporter. The Na(+)-dependent component of uptake is inhibited by 5-50 microM Hg2+ in a dose-dependent manner. p-Chloromercuribenzene sulfonic acid (p-CMBS) at high concentrations (500-1,000 microM) does not affect Na(+)-independent uptake but in the presence of Na+ depresses total uptake more than Na+ depletion alone, suggesting that Na+ enhances the binding of p-CMBS to both transporters. Spermidine and spermine compete with putrescine for uptake, but a variety of other organic bases and amino acids do not, indicating that polyamines are transported by mechanisms distinct from the transporters for those other compounds.


Subject(s)
Putrescine/metabolism , 4-Chloromercuribenzenesulfonate/pharmacology , Amiloride/pharmacology , Amino Acids/pharmacology , Animals , Biological Transport/drug effects , Cell Line , Cell Membrane/metabolism , Epithelium/metabolism , Kidney , Kinetics , Polyamines/pharmacology , Swine
20.
J Biol Chem ; 265(13): 7308-17, 1990 May 05.
Article in English | MEDLINE | ID: mdl-2110163

ABSTRACT

The amino acid sequence of the mitochondrial form of phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK-M) from the chicken was deduced from the 3571 nucleotide sequence of three overlapping cDNA clones. The derived protein sequence, which includes 607 amino acids of the mature enzyme and a leader sequence, was aligned with nine tryptic peptides of PEPCK-M and the primary sequence of the cytosolic form of PEPCK from chicken. Secondary structure predictions for the two PEPCK isozymes indicated similar packing elements of conserved, hydrophobic beta strands in the central core of the primary sequence. This core protein, which contained three GTP-binding consensus elements, was 80% identical in the two chicken isozymes, although the overall level of identity was only 63% for amino acids and 60% for nucleotides. The untranslated regions of the two cDNAs were dissimilar, although both mRNAs have potential for significant secondary structure. The PEPCK-M mRNA contained several G-C-rich regions which demonstrated free energies of formation in dyad symmetry programs up to -70 kcal/mol. The 1.6-kilobase (kb) 3'-untranslated region contained several repeat elements including one of 11 base pairs, which was present 30 times; but, a signal sequence for polyadenylation was not present. Each of the three PEPCK-M cDNA clones recognized two mRNAs of 4.2 and 3.4 kb in the livers and kidneys of starved or normally fed chickens. However, the level of these two related PEPCK-M mRNAs changed in response to cAMP treatment, with the larger mRNA predominant at 20 and 160 min and the 3.4-kb mRNA present at intermediate times. In contrast, the level of the 2.8-kb PEPCK-C mRNA increased dramatically upon addition of the cyclic nucleotide, particularly in the liver where it was not detected without cAMP induction. Thus, PEPCK-M and PEPCK-C, clearly represented the products of two distinct genes, which were distinguished by altered protein sequences and non-cross-hybridizing, differentially regulated mRNAs.


Subject(s)
DNA/genetics , Isoenzymes/genetics , Liver/enzymology , Mitochondria, Liver/enzymology , Phosphoenolpyruvate Carboxykinase (GTP)/genetics , Amino Acid Sequence , Animals , Base Sequence , Blotting, Northern , Chickens , Cytosol/enzymology , GTP-Binding Proteins/metabolism , Gene Library , Isoenzymes/metabolism , Molecular Sequence Data , Oligonucleotide Probes , Peptide Fragments/isolation & purification , Phosphoenolpyruvate Carboxykinase (GTP)/metabolism , RNA, Messenger/genetics , Restriction Mapping , Sequence Homology, Nucleic Acid
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