Alcohol dehydrogenase isozymes in the mouse: Genetic regulation, allelic variation among inbred strains and sex differences of liver and kidney A2 isozyme activity

Genetic analysis of a proposed cis-acting temporal locus ( Adh-3t ), which regulates alcohol dehydrogenase C₂ (ADH-C₂) acitivity in mouse epididymis extracts, among F₁ (ddN × BALB/c) × ddN male backcross progeny provided evidence for genetic distinctness between the structural ( Adh-3 ) and temporal ( Adh-3t ) loci on chromosome 3. Genetic analysis also confirmed the close, linkage of Adh-1 (encoding liver and kidney ADH-A₂) and Adh-3 (encoding stomach ADH-C₂) to within 0.3 centimorgans on the mouse genome. Evidence is presented for a proposed closely linked cis-acting temporal locus (designated Adh-1t ) for the A₂ isozyme (encoded by Adh-1 ) controlling the activity of this enzyme in mouse kidney extracts, but having no apparent affect on liver and intestine ADH-A₂ activities. An extensive survey of the distribution of Adh-1, Adh-3 and Adh-3t alleles among 65 strains of mice is reported — with the exception of two Japanese strains (ddN and KF), linkage disequilibrium between Adh-3 and Adh-3t was observed. Sex differences in mouse liver and kidney ADH-A₂ activities were observed, with male/female ratios of approximately 0.6 and 3 respectively for these tissue extracts.     

Ultrastructural changes in isolated rat hepatocytes exposed to different CCl4 concentrations

Ultrastructural data are presented on time-course changes in isolated rat hepatocyte suspensions exposed either to 1.2 or 1.8 mM CCl4 for up to 1 h. The subcellular changes at the lower concentration, but not the higher, are shown to closely parallel those reported to occur in rat hepatocytes following ingestion of CCl4.     

The effect of growth state on the activity of the protein kinase isoenzymes an increase in type II cyclic AMP-dependent protein kinase is correlated with growth arrest in G1 phase

Native polyacrylamide gels have been used to resolve protein kinase isoenzymes from cultured cells and the protein kinases have been identified by carrying out phosphorylation reactions in the gel. Following electrophoresis, the gels were incubated with histome and [γ-³²P]ATP. The gels were then thoroughly washed and dried down, and the protein kinases were located by autoradiography. Protein kinase activity as measured in the gel system was a linear function of cytosol protein concentration up to about 100 μg per channel and incorporation of ³²P into histone was time dependent. Three bands of protein kinase activity were resolved in cytosol samples from baby hamster kidney (BHK) fibroblasts. The band with the lowest relative mobility utilized histone IIA or casein equally well as substrate protein whereas bands 2 and 3 demonstrated a clear preference for histone. Bands 2 and 3 displayed a relative mobility in electrophoresis that was identical to that observed for cyclic AMP-dependent protein kinases I and II from rat liver. Treatment of cyctosol samples with cyclic AMP prior to electrophoresis resulted in the disappearance of cyclic AMP-dependent protein kinases from the gel profile. This method was employed to identify bands 2 and 3 as cyclic AMP-dependent protein kinases. The protein kinases in growth-arrested cells were compared with proliferating cells. We have observed a 3.5-fold increase in the activity of Type II protein kinase as the cells arrest growth in G₁ phase of the cell cycle. This increase in Type II is correlated with the increase in cells blocked in G₁ and a decrease in II Type activity appears to be an early event in permitting cells to leave G₁ and resume growth.     

An immunochemical analysis of the novel liver-restricted LDH activity from cichlid fishes (Cichlidae: Teleostei) confirms its non-orthology with piscine LDH-C

In an electrophoretic survey of lactate dehydrogenase (LDH) isozymes in neotropical cichlid fishes (Perciformes: Cichlidae) several species were discovered in which a cathodal liver-restricted isozyme is expressed along with the highly anodal eye-restricted isozyme (LDH-C₄) typically encountered in perciform fishes. Biochemical characterization of these two isozymes from the basketmouth cichlid, Acaronia nassa (Heckel), strongly suggested that they were non-orthologous and challenged the accepted view that eye- and liver-restricted LDH isozymes are alternative expressions of the same (LDH-C *) gene. In this study, antiserum raised against cypriniform (goldfish) liver-restricted LDH-C₄ failed to cross-react with the basketmouth liver-restricted analogue while effectively titrating the eye-restricted, anodal isozyme and, at higher titres, the LDH-B₄, heart-restricted isozyme from all cichlid species. Anti-serum raised against basketmouth muscle-restricted LDH-A₄ failed to titrate any of the eye- and liver-restricted isozymes. These data confirm the orthology of eye- and liver-restricted LDH isozymes in Cypriniform and Perciform fishes as originally proposed, suggest that the liver-restricted isozyme of cichlid fishes is non-orthologous and further raise the question of identity and evolutionary origin of this anomalous LDH activity.     

Allozyme variation and population structure of the very narrow endemic Seseli farrenyi (Apiaceae)

Analyses have been made of allozyme variation of the narrow endemic species Seseli farrenyi Molero & J. Pujadas (Apiaceae), which has only three known populations in Catalonia with a total of around 2000 individuals. All three populations were sampled and subjected to starch gel electrophoresis. Nine enzymes were resolved and 14 loci were interpreted. We detected high values of polymorphism ( P = 83.3%, A = 3.0, H _e = 0.297), far exceeding those expected for endemic species ( P = 26.3%, A = 1.39, H _e = 0.063). Genetic diversity was greater within populations than among populations, and the value of gene flow was very high ( Nm = 5.85). Most loci showed deviations from Hardy– Weinberg equilibrium, possibly due to the presence of subpopulations. The main threats to this species are human activities (tourism, fires), while natural threats are minimal due to its high genetic diversity. Finally, we propose some conservation measures which include both in situ and ex situ strategies. © 2000 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 138 , 305–314.     

Occurrence of Amidases in the Industrial Microbe Rhodococcus rhodochrous J1

Rhodococcus rhodochrous J1, of which the high-M_r nitrile hydratase has been used for the industrial manufacture of acrylamide from acrylonitrile, produced at least two amidases differing in substrate specificity, judging from the effects of various amides on amidase activity in this strain. These amidases seemed to be inducible enzymes depending on amide compounds.     

TK1与LDH联合检测对非霍奇金淋巴瘤鉴别诊断及疗效评估的作用

目的:探讨联合检测血清胸苷激酶1(TK1)与乳酸脱氢酶(LDH)水平在非霍奇金淋巴瘤(NHL)患者鉴别诊断及疗效监测中的临床意义。方法:收集2016年1月至2018年6月我院诊治的111例非霍奇金淋巴瘤的初诊患者血清标本,并选择50例正常人血清标本作为对照,采用免疫印迹增强发光法检测TK1浓度,比色法检测LDH浓度。所有患者随访至少1年,分析和比较惰性NHL与侵袭性NHL及各自四类分期之间血清TK1和LDH水平的差异,化疗后完全缓解、部分缓解与未缓解组LDH水平以及NHL患者中血清TK1和LDH的阳性率。结果:高度侵袭性NHL患者和侵袭性NHL患者血清TKI和LDH水平与惰性NHL患者相比显著增高(P0.05),但惰性NHL患者血清TK1和LDH水平与正常组之间差异无统计学意义(P0.05);Ⅲ、Ⅳ期侵袭性NHL患者血清TK1和LDH水平与Ⅰ、Ⅱ期患者相比显著增高(P0.05)。与化疗前相比,四次化疗后,完全缓解组NHL患者血清LDH水平下降21.05%,部分缓解组为16.66%,病情稳定组血清LDH水平升高至11.54%,三组NHL患者血清LDH水平比较差异具有统计学意义(P0.008),两组之间的差异均有统计学意义(P0.05)。结论:联合检测血清TK1和LDH水平对于NHL患者的鉴别诊断、疗效评估均具有重要参考价值。     

Mitochondrial free fatty acid β-oxidation supports oxidative phosphorylation and proliferation in cancer cells

Oxidative phosphorylation (OxPhos) is functional and sustains tumor proliferation in several cancer cell types. To establish whether mitochondrial β-oxidation of free fatty acids (FFAs) contributes to cancer OxPhos functioning, its protein contents and enzyme activities, as well as respiratory rates and electrical membrane potential (ΔΨm) driven by FFA oxidation were assessed in rat AS-30D hepatoma and liver (RLM) mitochondria. Higher protein contents (1.4–3 times) of β-oxidation (CPT1, SCAD) as well as proteins and enzyme activities (1.7–13-times) of Krebs cycle (KC: ICD, 2OGDH, PDH, ME, GA), and respiratory chain (RC: COX) were determined in hepatoma mitochondria vs. RLM. Although increased cholesterol content (9-times vs. RLM) was determined in the hepatoma mitochondrial membranes, FFAs and other NAD-linked substrates were oxidized faster (1.6–6.6 times) by hepatoma mitochondria than RLM, maintaining similar ΔΨm values. The contents of β-oxidation, KC and RC enzymes were also assessed in cells. The mitochondrial enzyme levels in human cervix cancer HeLa and AS-30D cells were higher than those observed in rat hepatocytes whereas in human breast cancer biopsies, CPT1 and SCAD contents were lower than in human breast normal tissue. The presence of CPT1 and SCAD in AS-30D mitochondria and HeLa cells correlated with an active FFA utilization in HeLa cells. Furthermore, the β-oxidation inhibitor perhexiline blocked FFA utilization, OxPhos and proliferation in HeLa and other cancer cells. In conclusion, functional mitochondria supported by FFA β-oxidation are essential for the accelerated cancer cell proliferation and hence anti-β-oxidation therapeutics appears as an alternative promising approach to deter malignant tumor growth.