Relationship between cellular ATP content and cellular functions of primary cultured rat hepatocytes in hypoxia

The importance of oxygen in maintaining the functional integrity of hepatocytes has been well established in a variety of experimental models, such as in vivo , perfused liver and isolated hepatocytes. However, one of the shortcomings of these systems is their short life span. Therefore, we have examined the effects of long-term hypoxia on cellular adenine nucleotide content and cellular functions, such as albumin production, urea production and DNA synthesis, in adult rat hepatocytes in primary culture. Hepatocytes were cultured at a density of 11 × 10⁴ and 5 × 10⁴ cells/0.18 mL per cm² for the study of albumin and urea production and DNA synthesis, respectively, at various oxygen tensions (20, 12, 8 and 5%) for 24 h. Cellular ATP content in cultured hepatocytes in hypoxia gradually declined, corresponding to the decrease in oxygen tension, and the cellular ATP level at 5% oxygen was approximately 20% of that at 20% oxygen. Albumin production also decreased in parallel with the decrease in cellular ATP content in cultured hepatocytes in hypoxia. However, even when cellular ATP content gradually declined corresponding with the decrease in oxygen tension in cultured hepatocytes in hypoxia, such as at 8 or 5% oxygen, urea production remained at a high level; in contrast, DNA synthesis was completely suppressed. These results suggest that the cellular ATP content decreases in cultured hepatocytes during long-term hypoxia in relation to oxygen tension and that the relationship between decreased ATP levels and liver function in cultured hepatocytes during hypoxia differs for albumin production, urea production and DNA synthesis.     

Cyclotron production of carrier-free 66Ga as a positron emitting label of albumin colloids for clinical use

A method for producing carrier free ⁶⁶Ga (T_1/2:9.4h; ⁺) by ⁴He bombardment of natural copper targets is presented. ⁶⁶Ga is formed by means of the ⁶³Cu (⁴He, n) ⁶⁶Ga reaction. Production yields are given in the 17.5 to 8 MeV ⁴He energy range. Chemical purification of ⁶⁶Ga from the copper target is described. The only radionuclidic impurity found in the final product was ⁶⁷Ga. Albumin colloids from commercially available kits designed for use with ^99mTc could easily be labeled with ⁶⁶Ga and employed for studies of the lymphatic system by positron emission tomography.     

Decreased antibody-dependent cellular cytotoxicity in minimal change nephrotic syndrome

Secondary IgG response to a tetanus toxoid booster and in vitro measurement of immunoglobulin synthesis, antibody-dependent cellular cytotoxicity (ADCC) and -interferon (IFN-) production were evaluated in 20 healthy controls and in 17 children with minimal change nephrotic syndrome (MCNS), during the acute nephrotic phase and 6 months after remission. Defective responses were observed in all but IFN- production during the acute nephrotic phase; these improved with disease remission. There was a significant correlation between decreases in vitro IgG production and ADCC reaction. These data indicate that defective antibody production is associated with decreased ADCC during the acute nephrotic phase of MCNS.     

2,4—D对甜橙幼果脱落及离区纤维素酶活性的抑制效应

暗柳橙(Citrus sinensis Osbeck)第一次生理落果期(3月下旬～4月中旬)的果实外植体脱落过程中,离区纤维素酶A、B的活性明显升高,第二次落果(4月下旬～5月中旬)期的果实脱落过程中,纤维素酶A的活性也显著提高,但纤维素酶B活性变化较小。细胞内、外纤维素酶的活性在外植体脱落过程中也不断升高。2,4—D能明显抑制胞内、外纤维素酶及纤维素酶A的活性,同时还能抑制外植体及非离休果实内乙烯的产生,但对纤维素酶B活性的影响无明显效应。田间喷布低浓度的2,4—D,明显地减少第一次生理落果,但对第二次落果的作用不如赤霉素。本文还讨论了果实的脱落、纤维素酶活性与乙烯之间的相互关系,及2,4—D的调控作用。并认为采用外植体的方法不失为研究脱落问题的有效手段。     

Hypohydration effects on thermoregulation during moderate exercise in the cold

Hyperosmotic hypovolemia impairs vasoconstriction during sedentary cold exposure. The purpose of this study was to determine whether hypohydration alters thermoregulation and cardiovascular responses to exercise in cold air. On four occasions, eight males [35.1 (2.7) years, 175.5 (3.1) cm, 73.3 (2.6) kg, 57.2 (2.6) ml kg^–1 min^–1 maximal oxygen uptake (O_2max), 19.6 (2.4)% fat] walked, in t-shirt, shorts, and shoes, at 50% O_2max, for 60 min in either a 4°C (Cold) or a 25°C (Temperate) environment in both hypohydrated state (HYPO, –4% body mass) and euhydrated state (EU). During exercise–cold stress, rectal temperature (T_re), mean weighted skin temperature, heart rate (HR), cardiac output (CO), and stroke volume (SV) were measured every 20 min. Mean weighted skin temperature values were not different between HYPO and EU but were lower (P<0.05) in Cold versus Temperate trials. T_re was not different (P>0.05) between HYPO–Cold and EU–Cold. CO and SV were not different within hydration states and were not different between Cold and Temperate trials (P<0.05). HR was not different between HYPO–Cold and EU–Cold. These data demonstrate that moderate intensity exercise in the cold while hypohydrated does not alter metabolic heat production, skin temperatures and heat loss, nor does it increase thermoregulatory and cardiovascular strain.     

IL-10-driven immunoglobulin production by B lymphocytes from IgA-deficient individuals correlates to infection proneness

In search for a possible explanation of the phenotypic heterogeneity in IgA deficiency, we studied the function of B cells from IgA-deficient (IgAd) individuals. Two groups of IgAd individuals, one frequently infected and one clinically apparently healthy, as well as normal controls, were studied. Peripheral blood mononuclear cells (PBMC) and B cells from IgAd individuals and controls were cultured with Staphylococcus aureus Cowan I strain and with anti-CD40 MoAb presented on the CD32-transfected fibroblast cell line in the presence of IL-10. In this experimental system PBMC and B cells from the infection-prone IgAd individuals produced only minute amounts of IgA. In contrast, PBMC and B cells from healthy IgAd subjects secreted significantly more IgA1 and IgA2 in comparison with infection-prone IgAd patients (P < 0.05). These data suggest that the abnormalities of B cell differentiation in IgAd could be of heterogeneous origin. Thus, whereas in healthy IgAd subjects IgA production may be efficiently up-regulated in vitro by addition of IL-10 to CD40-activated B cell culture, the corresponding B cell differentiation does not occur in infection-prone IgAd patients. These observations provide a conceptual framework for phenotypic heterogeneity in IgAd subjects.     

Predicting development of sustained unresponsiveness to milk oral immunotherapy using epitope-specific antibody binding profiles

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驱虫斑鸠菊注射液对小鼠免疫功能的影响

目的:探讨驱虫斑鸠菊对小鼠免疫功能的影响。方法:采用[3H]-TdR掺入法和脾细胞介导羊红细胞定量溶血分光光度法以及迟发型超敏反应试验等观察了驱虫斑鸠菊对小鼠兔疫功能的作用。结果:驱虫斑鸠菊在体内外均可以明显抑制ConA刺激的小鼠T淋巴细胞的增殖反应和LPS刺激的小鼠B淋巴细胞的增殖反应（P<0.01）;对小鼠胸腺指数和脾脏指数、绵羊红细胞（SRBC）诱导的正常小鼠脾抗体细胞生成反应以及小鼠皮肤迟发型超敏反应都显示出明显的抑制作用,而且上述抑制作用与药物浓度有一定的剂量效应关系。结论:驱虫斑鸠菊对机体体液免疫、细胞免疫都有明显的抑制作用。     

Mechanical and energy characteristics during shortening in isolated type-1 muscle fibres from Xenopus laevis studied at maximal and submaximal activation

 The mechanical and energy characteristics of isolated fast-twitch muscle fibres (type 1) of Xenopus laevis in isometric- and isovelocity contractions were measured at 20°C. The fibres were stimulated at either 60 Hz or 20 Hz to produce contractions at different levels of activation. The high stimulation frequency gave fused contractions, while at the low stimulation frequency tension fluctuated. When maximum isometric force had been reached, the fibres were shortened by 10% of the fibre length at different velocities. At 60 Hz stimulation during shortening the rate of heat production increased above the isometric rate of heat production. At 20 Hz stimulation during shortening, however, the rate of heat production was not different from the isometric rate of heat production. Mechanical efficiency was the same at the high and low level of activation. The actomyosin efficiency (i.e. the mechanical efficiency corrected for ”activation heat”) was highest at the low level of activation. We conclude that in fast-twitch muscle fibres from X. laevis, actomyosin efficiency is highest for partially activated muscle. From a comparison of the present results with those obtained from a study of slow-twitch muscle fibres presented earlier, it is concluded that fast-twitch muscle fibres are less efficient than slow-twitch muscle fibres. Received after revision: 13 May 1997 / Accepted: 1 July 1997     

Rapid microassays of phagocytosis, bacterial killing, superoxide and hydrogen peroxide production by human neutrophils in vitro

Simple, rapid microassays for simultaneous measurement of phagocytosis, bacterial killing, superoxide and hydrogen peroxide production by human neutrophils in vitro are described. All assays employ 96-well flat bottom tissue culture plates which were incubated on a microtitre plate shaker at 37 degrees C. The separate evaluation of ingestion and intracellular killing of E. coli and S. aureus was based on the incorporation of [3H]uridine into viable extracellular bacteria. There was good correlation between plate counts of viable bacteria and amount of radiolabel incorporation. Phagocytosis and killing can be measured in a maximum of 100 microliter reaction mixture, requiring only 2.5 X 10(5) neutrophils per test and the assay is complete within 60 min. Assay of superoxide production by stimulated neutrophils was based on superoxide-dependent reduction of ferricytochrome c as measured spectrophotometrically at 550 nm in wells of tissue culture plates containing 150 microliter of reaction mixture. The assay requires only 1.25 X 10(5) neutrophils per test and is complete within 50 min. Quantitation of hydrogen peroxide was based on horseradish peroxidase-dependent oxidation of phenol red. The technique is as for superoxide detection except that the reaction must be terminated by the addition of 1 M NaOH at the desired time intervals. None of the assays require sampling during the incubation period. The principal advantages of the described techniques are increased simplicity and speed, requirement of low numbers of neutrophils and applicability to analysis of large number of samples in parallel.