Peptidylarginine deiminase in rat and mouse hemopoietic cells

Summary Peptidylarginine (protein-L-arginine) deiminase activities have been demonstrated in extracts of rat and mouse peritoneal macrophages, bone marrow cells, splenic adherent cells, neutrophils, and mouse monocyte/macrophage cell lines. The enzyme in these cells is indistinguishable from the skeletal muscle enzyme with respect to immunochemical properties.     

Cytogenetic evidence for hemizygosity at the thymidine kinase locus in P388 mouse lymphoma cells

A detailed cytogenetic investigation was carried out on P388 mouse lymphoma cells. The cells have a mean chromosome number of 36.86 with a mode and median of 37 chromosomes. G-banding analysis of 12 spreads revealed a total of 15 marker chromosomes with chromosome 11, the determinant of thymidine kinase, being present only in single copy per cell. It is therefore concluded that the P388 cell line is hemizygous at the thymidine kinase locus. Thymidine kinase activities were assayed in P388 cells and two other malignant cell lines, clone 707 Friend mouse leukaemia cells and L5178Y mouse lymphoma cells. No clear relationship was observed between enzyme activity and gene dosage.     

茶多酚对裸鼠喉鳞癌的抑制作用及机制研究

目的观察萘多酚对裸鼠喉鳞癌的抑制作用，并初步探讨其机制。方法将接种Hep22细胞后的裸鼠随机分为4组，其中对照组8只，每天给予生理盐水1ml灌胃；茶多酚高、中、低剂量组各8只，每天分莉给予茶多酚80、160、320mg／kg。20d后处死。分离肿瘤测量其体积，real—time RT—PCR法检测肿癌组织中Src、HK Ⅱ mRNA的表这。己糖激酶活性测定试剂盒测定肿瘤组织中己糖激酶活性。结果茶多酚能硬显降低肿瘤体积，降低肿瘤组织中Src、HK Ⅱ mRNA的表达和己糖激酶活性。结论茶多酚能抑制裸鼠喉鳞癌的生长，其机制可能与抑制肿瘤糖酵解有关。     

Cell cycle patterns of thymidylate synthetase and 5,10-methylenetetrahydrofolate polyglutamates in cultured mouse hepatoma cells

Summary The regulation of thymidylate synthetase activity was investigated throughout the first cell cycle after release from an isoleucine block in synchronous cultures of mouse hepatoma (Hepa) cells. Activity in cell extracts increased with the onset of S phase and the increased activity was attributed to a parallel increase in enzyme concentration as determined by titration with tritiated fluorodeoxyuridylate. The polyglutamate chain length of reduced folate cofactors, which could also influence thymidylate synthetase activity, was unchanged.Acknowledgments. Supported by grant CA 22754, awarded by the National Cancer Institute, DHEW.     

Substrate induced alterations in tryptophan pyrrolase activity in two mouse strains

Summary Total hepatic L-tryptophan 2,3-dioxygenase activity was studied in 2 mouse strains receiving i.p. injections of L-tryptophan. After a single injection, enzyme activity was increased in albino but not pigmented mice. After 3 injections, enzyme activity was reduced in both strains. This work was partially supported by grant No. 3726-22 from the Office of Research and Sponsored Programs, Wichita State University.     

Sandwich enzyme immunoassay of mouse mammary tumor virus

Summary A highly sensitive sandwich enzyme immunoassay for the mouse mammary tumor virus (MMTV) is described. The assay can detect 3 ng/ml of MMTV. The enzyme used is ß-D-galactosidase fromEscherichia coli and the solid phase used is a piece of silicon rubber.     

Adenyl cyclase activity of mouse liver membranes after incubation with endotoxin and epinephrine.

Adenyl cyclase activity in isolated mouse liver cell membranes was stimulated two-fold by endotoxin. Furthermore, endotoxin inhibited epinephrine induction of adenyl cyclase activity, apparently through interruption of the phospholipid moiety of the enzyme complex.     

Adenyl cyclase activity of mouse liver membranes after incubation with endotoxin and epinephrine

Summary Adenyl cyclase activity in isolated mouse liver cell membranes was stimulated two-fold by endotoxin. Furthermore, endotoxin inhibited epinephrine induction of adenyl cyclase activity, apparently through interruption of the phospholipid moiety of the enzyme complex.     

Cross-reactive monoclonal antibodies to alcohol dehydrogenases

Summary Three anti-horse liver alcohol dehydrogenase (HLADH) monoclonal antibodies are described. Two are specific for ADH and cross-react with class I and II enzymes from mouse, horse and Chinese hamster. They are specific for the native enzyme but do not inhibit enzyme activity except when combined at high concentration. The third antibody was isolated as a response to rabbit metallothionein. It binds metalloproteins and inhibits ADH activity.     

Identification of type A monoamine oxidase in mouse and rabbit liver mitochondria

Summary Type A monoamine oxidase was identified in liver mitochondria of mouse and rabbit. 5-Hydroxytryptamine was a common substrate for type A and type B monoamine oxidase in these enzyme preparations where its concentration was 1.0 mM.     

Cross-reactive monoclonal antibodies to alcohol dehydrogenases

Three anti-horse liver alcohol dehydrogenase (HLADH) monoclonal antibodies are described. Two are specific for ADH and cross-react with class I and II enzymes from mouse, horse and Chinese hamster. They are specific for the native enzyme but do not inhibit enzyme activity except when combined at high concentration. The third antibody was isolated as a response to rabbit metallothionein. It binds metalloproteins and inhibits ADH activity.     

The effect of levamisole on phosphodiesterase activity.

Phosphodiesterase activity of mouse liver homogenates was estimated in presence and absence of levamisole. The enzyme activity was 1394 and 1399 nmoles/mg protein/30 min respectively. Our data show that levamisole does not affect the phosphodiesterase activity.     

The effect of levamisole on phosphodiesterase activity

Summary Phosphodiesterase activity of mouse liver homogenates was estimated in presence and absence of levamisole. The enzyme activity was 1394 and 1399 nmoles/mg protein/30 min respectively. Our data show that levamisole does not affect the phosphodiesterase activity.     

IgM memory B cells: a mouse/human paradox

Humoral memory is maintained by two types of persistent cells, memory B cells and plasma cells, which have different phenotypes and functions. Long-lived plasma cells can survive for a lifespan within a complex niche in the bone marrow and provide continuous protective serum antibody levels. Memory B cells reside in secondary lymphoid organs, where they can be rapidly mobilized upon a new antigenic encounter. Surface IgG has long been taken as a surrogate marker for memory in the mouse. Recently, however, we have brought evidence for a long-lived IgM memory B cell population in the mouse, while we have also argued that, in humans, these same cells are not classical memory B cells but marginal zone (MZ) B cells which, as opposed to their mouse MZ counterpart, recirculate and carry a mutated B cell receptor. In this review, we will discuss these apparently paradoxical results.     

Secificity of mouse endocervical cell response progesterone: presence of receptors]

The mouse cervical cell response to progesterone, corticosterone, and androgens was studied in vitro and comparatively by grafts on males. Contrary to the exocervical cells which responded more or less to the three steroid hormones, the endocervical cells responded exclusively to progesterone even in an estrogen free system. This result suggests the existence in the mouse endocervical cells of specific receptors to progesterone and indicates that the squamons cells of the uterine cervix have a different response to steroid hormones depending on where these cells are located.     

Steroid metabolism by mouse preimplantation embryos in vitro.

Mouse preimplantation embryos were incubated with radioactive pregnenolone, progesterone or dehydroepiandrosterone for various periods of time. These substrates were not converted to metabolites even after incubation of 120 h. We suggest that preimplantation mouse embryo does not possess enzyme activities for steroid metabolism.     

Tissue transglutaminase inhibits the TRPV5-dependent calcium transport in an N-glycosylation-dependent manner

Tissue transglutaminase (tTG) is a multifunctional Ca²⁺-dependent enzyme, catalyzing protein crosslinking. The transient receptor potential vanilloid (TRPV) family of cation channels was recently shown to contribute to the regulation of TG activities in keratinocytes and hence skin barrier formation. In kidney, where active transcellular Ca²⁺ transport via TRPV5 predominates, the potential effect of tTG remains unknown. A multitude of factors regulate TRPV5, many secreted into the pro-urine and acting from the extracellular side. We detected tTG in mouse urine and in the apical medium of polarized cultures of rabbit connecting tubule and cortical collecting duct (CNT/CCD) cells. Extracellular application of tTG significantly reduced TRPV5 activity in human embryonic kidney cells transiently expressing the channel. Similarly, a strong inhibition of transepithelial Ca²⁺ transport was observed after apical application of purified tTG to polarized rabbit CNT/CCD cells. Furthermore, tTG promoted the aggregation of the plasma membrane-associated fraction of TRPV5. Using patch clamp analysis, we observed a reduction in the pore diameter after tTG treatment, suggesting distinct structural changes in TRPV5 upon crosslinking by tTG. As N-linked glycosylation of TRPV5 is a key step in regulating channel function, we determined the effect of tTG in the N-glycosylation-deficient TRPV5 mutant. In the absence of N-linked glycosylation, TRPV5 was insensitive to tTG. Taken together, these observations imply that tTG is a novel extracellular enzyme inhibiting the activity of TRPV5. The inhibition of TRPV5 occurs in an N-glycosylation-dependent manner, signifying a common final pathway by which distinct extracellular factors regulate channel activity.     

Possible target of Abelson virus phosphokinase in cell transformation

By fusing interphase cells to cells undergoing mitosis, the interphase chromosomes can be visualized. When analyzed in this way, chromosomes of normal mouse cells show characteristic undercondensed centromeric regions. We have found that the centromeric regions of chromosomes from Abelson virus-transformed cells are fully condensed. Abelson virus transforms mouse cells by introducing into them a virally encoded phosphokinase that is expressed constitutively. Thus, we propose that the condensation of centromeric chromatin is a result of overphosphorylation by the Abelson virus phosphokinase, and that the centromeric region is the relevant target of overphosphorylation in transformed cell growth.     

Rapid purification of lactate dehydrogenase X from mouse testes by two steps of affinity chromatography on oxamate-sepharose.

At concentrations of 200 muM NADH and 0.5 M NaCl LDH-X is separated from the other LDH isozymes of mouse testes on oxamate-sepharose. In a second step LDH-X is bound to the same matrix at lower NADH and NaCl concentrations and the pure enzyme can subsequently be eluted.     

«Affinity Labelling» des cholinergischen Rezeptors der motorischen Endplatte mit Diazo-Acetylcholin

Summary Diazoacetylcholinebromide was used for affinity labelling of the cholinergic receptors of endplates in the mouse diaphragm. Under irradiation by light (Hg-high-pressure-lamp), irreversible depolarizing block of neuromuscular transmission developed. Acetylcholinesterase was influenced only by much higher concentrations, which demonstrates different localizations of the active enzyme center and the cholinergic receptor.