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1.
The plasma-lymph exchange of two lactate dehydrogenase (LDH) isozymes (4.2 nM) of different molecular charge between plasma (CP) and lung lymph (CL), their initial clearances across the capillary wall, and their extravascular distribution volumes were studied in dog lungs at control and increased left atrial pressures (Pla = 23.3 +/- 2.1 cmH2O). The anionic LDH 1 [isoelectric point (pI) = 5.0] exhibited a more rapid plasma decay compared with cationic LDH 5 (pI = 7.9) after bolus injection but was maintained relatively constant in several experiments using a constant infusion. The mean 10- to 12-min lung tissue clearances were 40% higher for LDH 5 than LDH 1 at control Pla and 120% higher at increased Pla. The CL/CP ratios of the anionic LDH 1 were consistently higher at 4-5 h after simultaneous injection than those of cationic LDH 5. However, the extravascular distribution volumes of LDH 5 were significantly higher in lymph equivalents than those of LDH 1 at 4 h in both Pla groups. Fixed negative charges located on endothelial cells, basement membrane, and interstitial matrix could explain the observed differences between LDH isozymes in CL/CP ratios, tissue uptake, and extra-vascular distribution volumes by the ionic interaction of LDH 5 with these negative change sites which would retard the plasma-lymph transport and enhance the extravascular distribution volume and initial clearance of these cationic macromolecules.  相似文献   

2.
The effects of alpha-naphthylthiourea (ANTU) on lung microvascular permeability to plasma proteins were studied in anesthetized open-chest dogs. Lymph flow (Jv) was recorded, and total protein in plasma and lymph was analyzed after cannulating a small prenodal lung lymphatic. The protocol involved four experimental periods. Period 1. During this base-line period the preparation stabilized and steady states were reached in Jv, lymph total protein, pulmonary arterial pressure (Ppa), and left atrial pressure (Pla). Period 2. Pla was increased to approximately 20 cmH2O and maintained at that level until Jv and protein measurements attained a new steady state. Period 3. After Pla was lowered to control levels, ANTU (5 mg/kg body wt) was infused intravenously and parameters were measured for 3 h. Period 4 Pla was again raised to the pre-ANTU levels of period 2 and maintained for an additional 2-3 h. The lymphatic total protein clearance increased 8.6-fold for an equivalent increase in pulmonary capillary pressure after ANTU. Vascular permeability was assessed by estimating the osmotic reflection coefficient (sigma d) for total protein at the pulmonary capillary membrane. Sigma d decreased from 0.65 to 0.40 following ANTU. From plasma protein fractions in four experiments, equivalent pore radii for the capillary membrane of 95 and 280 A were calculated after ANTU compared with 80 and 200 A for normal lung capillaries. In addition, extravascular lung water increased from 3.8 +/- 0.16 to 5.87 +/- 0.25 following ANTU and to 7.55 +/- 0.55 (g/g blood-free dry wt) when Pla was elevated with ANTU. The experimental design allowed quantitative assessment of the vascular permeability increase after ANTU by use of lymph protein fluxes that had minimal errors due to changes in surface area or lymph contamination from nonpulmonary structures.  相似文献   

3.
D Y Cai  M Tien 《Biochemistry》1990,29(8):2085-2091
The oxycomplexes (compound III, oxyperoxidase) of two lignin peroxidase isozymes, H1 (pI = 4.7) and H8 (pI = 3.5), were characterized in the present study. After generation of the ferroperoxidase by photochemical reduction with deazoflavin in the presence of EDTA, the oxycomplex is formed by mixing ferroperoxidase with O2. The oxycomplex of isozyme H8 is very stable, with an autoxidation rate at 25 degrees C too slow to measure at pH 3.5 or 7.0. In contrast, the oxycomplex of isozyme H1 has a half-life of 52 min at pH 4.5 and 29 min at pH 7.5 at 25 degrees C. The decay of isozyme H1 oxycomplex follows a single exponential. The half-lives of lignin peroxidase oxycomplexes are much longer than those observed with other peroxidases. The binding of O2 to ferroperoxidase to form the oxycomplex was studied by stopped-flow methods. At 20 degrees C, the second-order rate constants for O2 binding are 2.3 X 10(5) and 8.9 X 10(5) M-1 s-1 for isozyme H1 and 6.2 X 10(4) and 3.5 X 10(5) M-1 s-1 for isozyme H8 at pH 3.6 and pH 6.8, respectively. The dissociation rate constants for the oxycomplex of isozyme H1 (3.8 Z 10(-3) s-1) and isozyme H8 (1.0 X 10(-3) s-1) were measured at pH 3.6 by CO trapping. Thus, the equilibrium constants (K, calculated from kon/koff) for both isozymes H1 (7.0 X 10(7) M-1) and H8 (6.2 X 10(7) M-1) are higher than that of myoglobin (1.9 Z 10(6) M-1).(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

4.
We measured the regional distribution of pulmonary extravascular and interstitial water to examine the possibility that regional differences in microvascular pressure or tissue stress may cause regional differences in lung water. We placed chloralose-anesthetized dogs in an upright (n = 6) or supine (n = 7) position for 180 min. We injected 51Cr-labeled EDTA to equilibrate to the extracellular space and 125I-labeled albumin to equilibrate with plasma. At the end of the experiment, the lungs were removed, passively drained of blood, and inflated before rapid freezing. Lungs were divided into horizontal slices, and extravascular, interstitial, and plasma water, red cell volume, and dry lung weight were determined for each slice. We found that regional extravascular and interstitial water were constant throughout the lungs in both groups and that there were no significant differences between upright and supine dogs. There were no significant differences in hematocrit between slices. We conclude that gravity and body position have no measurable effect on either the total size of the extravascular and interstitial compartments or their regional distribution.  相似文献   

5.
中华姬鼠与大林姬鼠的同工酶差异   总被引:1,自引:0,他引:1  
傅必谦  李举怀 《兽类学报》1998,18(4):304-310
中华姬鼠(Apodemusdraco)和大林姬鼠(Apodemuspeninsulae)是形态学上十分相似的两种鼠类。为了对两种姬鼠的分类提供生物化学方面的依据,采用聚丙烯酰胺凝胶等电聚焦电泳方法比较和分析了两种姬鼠的LDH同工酶、EST同工酶和SOD同工酶的差异。结果表明,两种姬鼠的LDH同工酶酶谱基本相似,而EST同工酶和SOD同工酶酶谱则存在明显的种间差异。根据EST同工酶A2带的有无和SOD同工酶主带等电点的差别,能将两种姬鼠很容易区分开来。  相似文献   

6.
黑爪异鲵LDH同工酶谱系的研究   总被引:9,自引:0,他引:9  
徐晋麟  马学海 《动物学报》1996,42(2):182-188
采用聚丙烯酰胺凝胶电泳对黑爪异鲵12种组织的LDH同工酶谱系进行了研究,酶谱呈5带型,但在雄性肝中出现了LDH-C带,在精巢中未发现,相对迁移率为B>A>C。 经比较分析,其ldh-C基因表达的组织特异性和性别特异性不仅证实黑爪异鲵是小鲵科中一个较原始的类群,且表明ldh-C基因这种表达调控方式在进化中为鱼类与鸟类。哺乳类之间的一种过渡类型。  相似文献   

7.
To examine whether urinary excretions of plasma proteins with molecular radii of 45-55 A and different isoelectric points such as IgG (pI = 7.4) and ceruloplasmin (pI = 4.4) increase selectively in normoalbuminuric type 2 diabetic patients, urinary albumin excretion rate (AER), renal clearances of IgG, ceruloplasmin and alpha2-macroglobulin, and creatinine clearance (Ccr) were studied in timed overnight urine samples of 36 diabetic outpatients and 16 control subjects. Furthermore, to examine effect of glycemic control on these urinary protein excretions, the same analysis was performed before and after glycemic control in 17 diabetic inpatients admitted for glycemic control. Renal clearances of IgG and ceruloplasmin were significantly higher in diabetic outpatients than in the control group, whereas AER and renal clearance of alpha2-macroglobulin did not differ. Glycemic control caused significant decreases in renal clearances of IgG and ceruloplasmin, accompanied with tendency for Ccr to decrease (p = 0.055). The present results, together with our previous finding of selectively increased urinary excretions of 45-55 A sized plasma proteins in parallel with enhanced glomerular filtration rate after acute protein loading, led us to conclude that enhanced intraglomerular hydraulic pressure may cause increases in clearances of IgG and ceruloplasmin, and that this change can be reversed by strict glycemic control in normoalbuminuric diabetic patients.  相似文献   

8.
The ontogeny of lactate dehydrogenase (LDH) isozymes in developing Mexican axolotl, A mbystoma mexicanum was investigated by thin-layer isoelectric focusing in polyacrylamide gel. The isoelectric points (pI values) of the isozymes were determined. The minor components generally remained masked during conventional electrophoresis, but became sharp as isofocusing progressed.
We identified in developing eggs and embryos five major LDH isozymes, which could also be traced in the ovarian eggs. All these isozymes, except LDH-1, consisted of one major and one minor component. Heterogeneity in axolotl LDH is reported for the first time. The separated isozymes had pI values from 5.24–6.60. Contrary to observations made by others, it was found that the anodal forms of LDH (pIs 5.24–5.80) were prominent throughout, while the remainder (pIs 6.16–6.60) gradually lost their stain ability.
It thus appears that isoelectric focusing is a possible method for the analysis of protein mixtures and can be successfully applied to problems of differentiation.  相似文献   

9.
To clarify the mechanism of aluminum (Al) toxicity and Al tolerance, we isolated a new clone (pAL201) from a tobacco cDNA library. Northern blot hybridization analysis indicated that the expression of pAL201 is induced by Al treatment and phosphate (P1) starvation. The complete cDNA sequence suggested that this clone encodes a moderately anionic peroxidase (EC 1.11.1.7). Analysis by isoelectric focussing indicated that a moderately anionic peroxidase (approximately pI 6.7) and two cationic peroxidases (pI 9.2 and 9.7) in the soluble fraction are activated by Al treatment and P1 starvation, while two moderately anionic isozymes are repressed by these stresses. We suppose that Al ion stress can control the activity of some peroxidase isozymes, one of which is probably induced by enhanced gene expression of pAL201. There is a possibility that some of these isozymes have some functions in Al ion stress.  相似文献   

10.
We investigated the effect of positive end-expiratory pressure (PEEP) on the extravascular thermal volume of the lung (ETV) determined by the thermal-dye technique in three canine models of pulmonary edema created by injection of alpha-naphthylthiourea (ANTU) or oleic acid (OA) into the pulmonary circulation or intrabronchial instillation of hydrochloric acid (HCl). ETV was determined before, during, and after ventilation with 14 cmH2O PEEP, and final ETV was compared with the extravascular lung mass (ELM) determined postmortem. Final ETV correctly estimated ELM in 12 animals with ANTU injury, ETV/ELM = 1.04 +/- 0.13, but underestimated after HCl injury (n = 5), ETV/ELM = 0.61 +/- 0.23, and OA injury (n = 6), ETV/ELM = 0.73 +/- 0.19. Whereas PEEP had no consistent effect on extravascular thermal volume in ANTU edema, there was a reversible increase in ETV during PEEP in animals with HCl or OA injury and underestimation of ELM. The increase in ETV during PEEP averaged 9.3 +/- 3.8 ml/kg (62 +/- 42%) over the mean of the pre- and post-PEEP values after HCl injury (P less than 0.01) and 6.7 +/- 4.4 ml/kg (47 +/- 35%) after OA injury (P less than 0.02). There was an inverse correlation between the change in ETV during PEEP and the ETV/ELM ratio for animals with HCl and OA injury (r = -0.94). We conclude that PEEP produces a reversible increase in ETV in some models of lung injury by allowing for distribution of thermal indicator through a larger fraction of the lung water and that this response may be useful to detect underestimation when gravimetric measurements are not available.  相似文献   

11.
1. It has been reported that DNase I can be highly purified from pancreas extract by affinity chromatography on a dDNA-Sepharose column under non-digestive conditions. In the present study, the adsorption-elution of other nucleases on the column under non-digestive conditions was studied. 2. All the seven kinds of nucleases tested were adsorbed when applied on a dDNA-Sepharose column under conditions which did not allow the enzymes to hydrolyze the DNA. The non-digestive conditions were as follows. i) For DNase II (pI=10.2), pH 3.0 in the presence of 50 mM sodium sulfate (inhibitor), ii) for micrococcal nuclease (pI=9.6), pH 4.0 in the absence of Ca2+ (activator), iii) for restriction endonucleases Eco RI (pI=5+1), Hind III (pI=5+1), and Bam HI (pI=5+1), pH 4.0 in the presence of 20% glycerol and 0.1% Neopeptone (stabilizers), and iv) for nucleases S1 (pI=5+1) and nuclease P1 (pI=4.5), pH 7.0. At the respective pH's, the enzymes other than nucleases S1 and P1 were cationic so as to exhibit electrostatic attraction to the anionic dDNA-Sepharose. Although S1 and P1 were anionic, they still adsorbed to the column. 3. All the adsorbed nucleases described above were eluted by a concentration gradient of KCl without changing pH. The ionic strengths required for elution were 0.19 for DNase II, 0.53 for micrococcal nuclease, 0.73 for Eco RI, 0.72 for Hind III, 0.37 for Bam HI, 0.17 for P1, and 0.13 for S1. The fact that the ionic strength required for the elution of DNase I (pI=5.0) was 0.39 at pH 4.0 indicates that the former five enzymes except DNase II can be chromatographed with almost the same or higher efficiency than DNase I, because the proteins adsorbed with no-specific affinity could be mostly eluted at lower ionic strength. On the other hand, the fact that nucleases P1 and S1 were adsorbed in spite of electrostatic repulsion suggests that these two enzymes can also be effectively chromatographed, especially when other cationic proteins are previously removed by an appropriate method such as adsorption to a typical cation exchanger.  相似文献   

12.
本文利用SOD和LDH在一块凝胶板上同时显色的优点,电泳分析黑线姬鼠(Apodemus agrarius)、褐家鼠(Rattus norvegicus),黄胸鼠(Rattus flavipectus)血浆SOD和LDH同工酶以及血浆蛋白成分。结果表明,LDH为有色区带,SOD为无色透明区带,而且SOD比LDH泳动快,两类区带互不重迭,以致3种鼠血浆中SOD和LDH的图谱清晰可辨。由于黑线姬鼠的SOD和LDH的泳动度不同于其余2种鼠,周此3种鼠的图谱各具特征。3种鼠血浆蛋白中的前清蛋白和铁传递蛋白区带数和泳动度亦具有明显差异,说明上述酶分子亚基和血浆蛋白分子的化学结构不同,成为种间差异在分子水平上典型的表现。  相似文献   

13.
Alpha‐naphthylthiourea (ANTU), a rodenticide induces lung toxicity. Chrysin a flavonoid possesses antioxidant, anti‐inflammatory, and antihypertensive potential. The aim of this study was to evaluate the efficacy of chrysin against ANTU‐induced pulmonary edema (PE) and pulmonary arterial hypertension (PAH) in laboratory rats. Sprague‐Dawley rats were used to induce PE (ANTU, 10 mg/kg, ip) and PAH (ANTU, 5 mg/kg, ip, 4 weeks). Animals were treated with chrysin (10, 20, and 40 mg/kg) and various biochemical, molecular, and histological parameters were evaluated. Acute administration of ANTU induces PE revealed by significant (P < 0.05) increase in relative lung weight, pleural effusion volume, lung edema, bronchoalveolar lavage fluid cell counts, total protein, 5‐hydroxytryptamine (5‐HT), lactate dehydrogenase (LDH), and γ‐glutamyl transferase (GGT), whereas pretreatment with chrysin (20 and 40 mg/kg, ip) significantly (P < 0.05) attenuated these ANTU‐induced biochemical and histological alterations. Repeated administration of ANTU caused induction of PAH evaluated by significant (P < 0.05) alterations in electrocardiographic, hemodynamic changes, and left ventricular function, whereas chrysin (20 and 40 mg/kg, p.o.) treatment significantly (P < 0.05) attenuated these alterations. ANTU‐induced hematological and serum biochemical (aspartate transaminase, alanine transaminase, LDH, and creatinine kinase MB) alterations were significantly (P < 0.05) inhibited by chrysin. It also significantly (P < 0.05) decreased elevated levels of oxido‐nitrosative stress in the right ventricle (RV) and lung. Chrysin significantly (P < 0.05) attenuated downregulated endothelial nitric oxide synthase and upregulated vascular endothelial growth factor messenger RNA and protein expressions both in the RV and pulmonary artery. Chrysin inhibited ANTU‐induced PE and PAH via modulation of inflammatory responses (5‐HT, LDH, and GGT), oxido‐nitrosative stress, and VEGF and eNOs levels.  相似文献   

14.
An increase in exocarp peroxidase activity was observed in fruit at 5 to 30 days post pollination (DPP), and decreased at 40 and 50 DPP. Total peroxidase activity of the mesocarp was significantly lower than the exocarp in all developmental stages. Mesocarp peroxidase activity decreased consecutively from outer, to middle and, to inner tissue at every developmental stage. Total activity in the mesocarp peaked at 20 DPP. Native-PAGE of exocarp tissue showed at least two cathodic (basic) peroxidases and two anionic (acidic) peroxidases. The number of isozymes was greatest and bands most intense at 30 DPP. IEF-PAGE of the 5 to 50 DPP fruit exocarp showed at least 8 peroxidase isozymes (pI 4.6 to 9.6). Anion exchange chromatography showed only one peak of anionic peroxidase activity that was not evident until 15 DPP. This peak was greatest at 30 DPP and declined at 40 and 50 DPP. Cationic peroxidase isozymes appeared to be the predominant and most intense isoforms throughout fruit development. The changes in peroxidase activity corresponded to fruit formation and may be associated with susceptibility to fruit rot.  相似文献   

15.
We assessed pulmonary endothelial and epithelial permeability and lung lymph flow in nine adult sheep under base-line conditions and after resuscitation from profound hemorrhagic shock. Animals were mechanically ventilated and maintained on 1% halothane anesthesia while aortic pressure was held at 40 Torr for 3 h. Systemic heparin was not used. After reinfusion of shed blood, sheep recovered from anesthesia and we measured lung lymph flow (QL), lymph-to-plasma concentration ratio for proteins, and time taken to reach half-equilibrium concentration of intravenous tracer albumin in lymph (t1/2). Twenty-four hours after bolus injection of radio-albumin we lavaged subsegments of the right upper lobe and determined fractional equilibration of the tracer in the alveolar luminal-lining layer. In each sheep we had measured these parameters 7 days earlier under base-line conditions. Animals were killed, and the lungs were used for gravimetric determination of extravascular lung water (gravimetric extravascular lung water-to-dry weight ratio) 24 h after resuscitation from shock. Pulmonary endothelial injury after resuscitation was evidenced by marked increase in QL, without fall in lymph-to-plasma ratio. Time taken to reach half-equilibrium concentration fell from 169 +/- 47 (SD) min in base-line studies to 53 +/- 33 min after shock. There was no evidence of lung epithelial injury. Gravimetric extravascular lung water-to-dry weight ratio was significantly increased in these animals killed 24 h after resuscitation (4.94 +/- 0.29) compared with values in our laboratory controls (4.13 +/- 0.09, mean +/- SD). These data demonstrate a loss of lung endothelial integrity in sheep after resuscitation from profound hemorrhagic shock.  相似文献   

16.
Vascular permeability and late radiation fibrosis in mouse lung   总被引:1,自引:0,他引:1  
It has been suggested that fibrosis which develops after irradiation is caused by increases in vascular permeability. Plasma proteins leak into irradiated tissue where fibrinogen may be converted into fibrin which is gradually replaced by fibrous tissue. Vascular and fibrotic changes in mouse lung were investigated after X irradiation of the right hemithorax. Blood volume and accumulation of extravascular proteins were measured using indium (111In)-labeled red cells, iodinated (131I) albumin, and iodinated (125I) fibrinogen. Tracers were injected 1-47 weeks after irradiation and lungs were excised 24 or 96 hr later to determine radioactivity. The amount of collagen was estimated by measuring the hydroxyproline content. During the first few months after X rays, lung blood volume decreased to a plateau which depended on radiation dose (10-25 Gy). Small increases in extravascular albumin and fibrinogen occurred at 1-12 weeks after 10-25 Gy. Subsequently, protein returned to normal after 10 Gy, remained elevated after 15 Gy, and increased after 20 and 25 Gy. Hydroxyproline per gram of dry irradiated lung was increased at 18 weeks after 15-25 Gy. Subsequently it showed little change although both total hydroxyproline content and dry weight decreased after 20 and 25 Gy. Support for the hypothesis was that hydroxyproline per gram only increased after X-ray doses which caused marked extravasation of protein. There was no evidence, however, for deposition of 125I-fibrin or for a gradual increase in fibrosis corresponding to the prolonged excess of extravascular protein.  相似文献   

17.
18.
Synthesis of COP (prostaglandins; PG and thromboxanes; Tx) from exogenous and endogenous arachidonic acid (AA) was studied in isolated perfused lungs from rats treated in vivo with a single dose of alpha-naphthylthiourea (ANTU; 10mg/kg;). Lung dry:wet weight ratios showed changes characteristic of oedema between 6 and 16h after ANTU. Bioassay of COP showed that COP synthesis from exogenous AA was raised above control values in lungs from rats treated with ANTU, reaching a maximum at 16h after treatment. By radioimmunoassay, the major increase was in 6-oxo-PGF1 alpha, with lesser effects on PGE2 and PGF2 alpha levels. Synthesis of bioassayable COP from endogenous AA induced by the calcium ionophore A23187 was increased as early as 2h after ANTU treatment and remained elevated up to 70h. In lungs 28h after ANTU, 6-oxo-PGF1 alpha release was greater than in normal lungs. These results show that in this model of pulmonary oedema, the potential for COP synthesis was increased. From the time course of this effect, increased COP synthesis was probably a response to the initial damage rather than a cause of the oedema.  相似文献   

19.
In the present study, we investigated the effects of the nitric oxide (NO) synthase inhibitor N(G)-nitro-L-arginine-methyl ester (L-NAME) on tissue injury or cytotoxicity caused by endotoxin challenge by assaying lactate dehydrogenase (LDH) isozymes and cell viability in J774A.1 cells. In mice treated with L-NAME (10 mg kg(-1), i.v.), the activity of LDH in serum 18 h after endotoxin (6 mg kg(-1), i.p.) injection was not significantly different from that in mice treated with endotoxin alone. Mice injected with endotoxin exhibited leakage of LDH isozymes 3 and 5, but L-NAME did not protect against endotoxin-induced acute leakage of LDH isozymes. Treatment with L-NAME (10-1000 microM) significantly inhibited NO generation by endotoxin (1 microg ml(-1))-activated J774A.1 cells. However, L-NAME (10-1000 microM) did not affect endotoxin-induced cytotoxicity in J774A.1 cells. These findings suggested that endotoxin-induced NO formation may not contribute to tissue injury or cytotoxicity caused by endotoxin.  相似文献   

20.
The dynamics of changes in the lactate dehydrogenase (LDH) spectra during organogenesis in CBA mice has been studied by means of ultramicroelectrophoresis. The embryonic period of development is characterized by the predominance of cathodic isozymes (LDH-5 and LDH-4) in all the organs under study. The increase of anodic isozymes (LDH-1 and LDH-2) takes place in the heart, kidneys and brain as the development proceeds. The first reliable differences in the LDH spectra of different organs appear on the 11th day of embryogenesis. On the basis of comparison with the help of criterion gamma, the LDH spectra of the organs under study can be divided into two groups: I--heart, lungs, kidneys and brain (tendency towards the increase of H-subunits) and II--intestine, liver and muscles (tendency towards the increase of of M-subunits). The LDH spectra of adult animals are divided into 4 distinct groups: I--heart and kidneys, II--brain, III--lungs and muscles, IV--liver.  相似文献   

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