Thyroxine-induced redistribution of creatine kinase isoenzymes in rat cardiomyocyte cultures

The present study demonstrates a change occurring in the creatine-kinase isoenzyme profile of cardiomyocyte cultures induced by a chronic administration of excessive amounts of thyroid hormones (TH). This change is manifested by an increased level of the CK-BB isoenzyme, generally at the expense of CK-MM isoenzyme. The elevation of CK-BB is probably a result of a specific effect of TH through activation of gene expression, rather than a contribution of an increased number of non-myocardial cells. The implications of these results in the diagnosis of heart failures are discussed.     

Influence of castration and testosterone replacement on hypothalamic tyrosine hydroxylase activity in the rat.

Hypothalamic tyrosine hydroxylase (TH) activity of castrate rats is modulated by testosterone propionate (TP) in vivo. Kinetic studies revealed that both Vmax and Km were virtually unaltered for substrate tyrosine in the presence of an excess of DMPH4 cofactor. TP replacement to castrate rats increased the Km for added DMPH4 cofactor, while Vmax decreased. These results suggest that TP decreases TH activity of castrate rats by inhibiting the enzyme-reduced pteridine cofactor complex.     

Hypothalamic tyrosine hydroxylase activity, plasma gonadotropin and prolactin levels after aminooxyacetic acid in ovariectomized rats

Plasma concentrations of gonadotropin, prolactin and hypothalamic tyrosine hydroxylase (TH) activity were measured in ovariectomized rats treated with aminooxyacetic acid (AOAA), a drug which elevates brain GABA levels. Hypothalamic TH activity was significantly increased with a significant decrease in prolactin (Prl) release. Plasma levels of gonadotropins were not modified by AOAA. These results support an inhibitory action of GABA on Prl release possibly mediated through hypothalamic dopamine.     

Hypothalamic tyrosine hydroxylase activity,plasma gonadotropin and prolactin levels after aminooxyacetic acid in ovariectomized rats

Summary Plasma concentrations of gonadotropin, prolactin and hypothalamic tyrosine hydroxylase (TH) activity were measured in ovariectomized rats treated with aminooxyacetic acid (AOAA), a drug which elevates brain GABA levels. Hypothalamic TH activity was significantly increased with a significant decrease in prolactin (Prl) release. Plasma levels of gonadotropins were not modified by AOAA. These results support an inhibitory action of GABA on Prl release possibly mediated through hypothalamic dopamine.Supported by grants from Indian Council of Medical Research (ICMR), New Delhi. RIA kits for the estimation of LH, FSH and Prl were kindly supplied by Dr A.F. Parlow, NIAMDD-NIH, Bethesda, Maryland, USA. GNB is a UGC research fellow.     

Tyrosine hydroxylase activity and tyrosine titers during cockroach metamorphosis

Summary Tyrosine hydroxylation (TH) to dopa in vivo in nymphs and pharate adults ofPeriplaneta americana was very low, but increased over 30-fold beginning at ecdysis. Free tyrosine increased in pharate, adults and peaked at ecdysis, whereas TH activity peaked 6 h later. Both TH activity and tyrosine titers declined the next 24 h as the cuticle tanned. TH appears to regulate tanning substrate biosynthesis in cockroaches.Contribution No. 81-294-j, Department of Entomology, Kansas Agricultural Experiment Station, Manhattan KS. Research supported in part by National Science Foundation grant No. PCM-8003859. I wish to thank Dr. M.A.J. Al-Izzi for assistance with the enzyme analyses.     

Influence of castration and testosterone replacement on hypothalamic tyrosine hydroxylase activity in the rat

Summary Hypothalamic tyrosine hydroxylase (TH) activity of castrate rats is modulated by testosterone propionate (TP) in vivo. Kinetic studies revealed that bothV _max andK _m were virtually unaltered for substrate tyrosine in the presence of an excess of DMPH₄ cofactor. TP replacement to castrate rats increased theK _m for added DMPH₄ cofactor, whileV _max decreased. These results suggest that TP decreases TH activity of castrate rats by inhibiting the enzymereduced pteridine cofactor complex.     

Changes in lactic dehydrogenase isoenzyme pattern in muscle of tumor-bearing mice

Summary The lactic dehydrogenase isoenzyme distribution shifts gradually toward the isoenzyme pattern of the tumor in mouse muscles located distally from a transplanted mammary carcinoma.This study was supported by a grant from the Minnesota Masonic Memorial Cancer Research Fund and, in part, by Grant No. 16-P-5681015-17 from the Rehabilitation Services Administration, U.S. Dept. of Health, Education and Welfare.     

Pyruvate kinase inhibited by L-cysteine as a marker of tumorigenic human urothelial cell lines

Summary It was found that a decrease in electrophoretic mobility of pyruvate kinase (PK) isoenzyme, and an increase of the sensitivity of this enzyme to L-cysteine, were markers of immortalization and tumorigenic properties, respectively, in human urothelial cell lines characterized by different grades of transformation (TGr) in vitro.     

Somatic cell hybrids producing inhibitors of melanotic melanoma tyrosine hydroxylase

Summary Splenic lymphocytes from BALB/c mice pre-immunized with purified tyrosine hydroxylase (TH) were fused with murine myeloma N/1 cells. Supernatants of only 2 from large number of cloned cell hybrids contained an inhibitor of TH.     

Adenosine triphosphatase ofAspergillus nidulans: Stimulation by aminoacids

Summary Ca²⁺-dependent ATPase ofAspergillus nidulans was found to be stimulated by aminoacids in vitro. Both histidine and arginine stimulated the enzyme more effectively than the aromatic aminoacids. Supplementation of the growth medium with basic or aromatic aminoacids increased the enzyme activity in vivo 2–6-fold. The enhanced activity observed in these cultures in vivo was not mediated through the synthesis of new isoenzyme, as observed in proteinenriched cultures, but appeared to be through the activation of enzyme activity.     

Pyruvate kinase inhibited by L-cysteine as a marker of tumorigenic human urothelial cell lines

It was found that a decrease in electrophoretic mobility of pyruvate kinase (PK) isoenzyme, and an increase of the sensitivity of this enzyme to L-cysteine, were markers of immortalization and tumorigenic properties, respectively, in human urothelial cell lines characterized by different grades of transformation (TGr) in vitro.     

A sensitive and inexpensive high-performance liquid chromatographic assay for tyrosine hydroxylase

We describe a highly sensitive assay method for tyrosine hydroxylase (TH) using high-performance liquid chromatography with amperometric determination. This assay method could be applicable to any tissues with low enzyme activity, such as rat cerebellum. We also describe the kinetic properties of TH in rat cerebral cortex.     

Isoenzyme patterns in androgenic,haploidDatura meteloides (Solanaceae)

Summary Leaf tissue isoenzyme patterns of androgenically derived, haploid plants ofDatura meteloides were compared with patterns from diploid parental plants. No isoenzyme patterns unique to the haploid palnts were detected among 5 enzymes studied.     

A sensitive and inexpensive high-performance liquid chromatographic assay for tyrosine hydroxylase

Summary We describe a highly sensitive assay method for tyrosine hydroxylase (TH) using high-performance liquid chromatography with amperometric determination. This assay method could be applicable to any tissues with low enzyme activity, such as rat cerebellum. We also describe the kinetic properties of TH in rat cerebral cortex.     

Ranbp2 haploinsufficiency mediates distinct cellular and biochemical phenotypes in brain and retinal dopaminergic and glia cells elicited by the Parkinsonian neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)

Many components and pathways transducing multifaceted and deleterious effects of stress stimuli remain ill-defined. The Ran-binding protein 2 (RanBP2) interactome modulates the expression of a range of clinical and cell-context-dependent manifestations upon a variety of stressors. We examined the role of Ranbp2 haploinsufficiency on cellular and metabolic manifestations linked to tyrosine-hydroxylase (TH(+)) dopaminergic neurons and glial cells of the brain and retina upon acute challenge to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a parkinsonian neurotoxin, which models facets of Parkinson disease. MPTP led to stronger akinetic parkinsonism and slower recovery in Ranbp2 (+/-) than wild-type mice without viability changes of brain TH(+)-neurons of either genotype, with the exception of transient nuclear atypia via changes in chromatin condensation of Ranbp2 (+/-) TH(+)-neurons. Conversely, the number of wild-type retinal TH(+)-amacrine neurons compared to Ranbp2 (+/-) underwent milder declines without apoptosis followed by stronger recoveries without neurogenesis. These phenotypes were accompanied by a stronger rise of EdU(+)-proliferative cells and non-proliferative gliosis of GFAP(+)-Müller cells in wild-type than Ranbp2 (+/-) that outlasted the MPTP-insult. Finally, MPTP-treated wild-type and Ranbp2 (+/-) mice present distinct metabolic footprints in the brain or selective regions thereof, such as striatum, that are supportive of RanBP2-mediated regulation of interdependent metabolic pathways of lysine, cholesterol, free-fatty acids, or their β-oxidation. These studies demonstrate contrasting gene-environment phenodeviances and roles of Ranbp2 between dopaminergic and glial cells of the brain and retina upon oxidative stress-elicited signaling and factors triggering a continuum of metabolic and cellular manifestations and proxies linked to oxidative stress, and chorioretinal and neurological disorders such as Parkinson.     

Recovery of renal lactate dehydrogenase (LDH) isoenzyme pattern after obstruction relief in experimental hydronephrosis

Summary The release of ureteral occlusion leads to a progressive recovery in LDH isoenzyme pattern with gradual increase of anodic fractions and decrease of middle and cathodic ones. Our findings demonstrate that the recovery is accomplished on the 10–14th day, in agreement with morphological and metabolic observation.This work was partially supported by research grant C.N.R. 730056004.     

Recovery of renal lactate dehydrogenase (LDH) isoenzyme pattern after obstruction relief in experimental hydronephrosis.

The release of ureteral occlusion leads to a progressive recovery in LDH isoenzyme pattern with gradual increase of anodic fractions and decrease of middle and cathodic ones. Our findings demonstrate that the recovery is accomplished on the 10-14th day, in agreement with morphological and metabolic observation.     

Immunohistochemical localization of carbonic anhydrase C in the rat central nervous system]

With obtention of specific antibodies against isoenzyme C of Rat carbonic anhydrase it was possible to realize an immunohistochemical localisation in central nervous system of Rat. Results showed that this enzyme was mainly localized in cytoplasm of oligodendroglial cells and in low concentration in myelin sheath.     

2-Methoxybenzoyl phosphate: a new substrate for continuous fluorimetric and spectrophotometric acyl phosphatase assays

A new aromatic acyl phosphate, 2-methoxybenzoyl phosphate, has been synthesized. The compound shows an intrinsic fluorescence; it displays an intense emission band at 390 nm upon excitation in the near UV region. This band practically disappears after hydrolysis of the product. On the other hand, the product displays differences in the near UV absorption spectra measured before and after hydrolysis. The at 301 nm is 2720 M^–1 cm^–1, a value that is 4.3-fold higher than that of benzoyl phosphate (the usual substrate for acylphosphatase assay) at 283 nm. The main kinetic parameters of three different acylphosphatase molecular forms (the muscular isoenzyme and two subtypes of the organ common isoenzyme) were determined using both benzoyl phosphate and 2-methoxybenzoyl phosphate as substrates, and then compared. These kinetic data and the UV absorption and fluorescence properties of 2-methoxybenzoyl phosphate sugest that this compound has better substrate features than benzoyl phosphate, and can be used for both high sensitivity continuous fluorimetric and UV absorption spectrophotometric assays of acylphosphatase.