Chronic overcrowding decreases cytoplasmic free calcium levels in T lymphocytes of aged CBA/CA mice

Intracellular calcium concentration is a sensitive marker of the homeostasis of living cells, and its increase is an essential step of T lymphocyte activation. Changes in the environment provoke an adaptive stress-response of the organism. In our present work we have investigated the effect of chronic overcrowding on resting and lectin-stimulated cytoplasmic free calcium concentration of splenic T lymphocytes from young and aged CBA/CA mice (50 animals total). The animals were kept under ‘normal’ (68 cm²/animal) or ‘overcrowded’ (22 cm²/animal) conditions for 3 months. Young animals showed no change in resting and stimulated calcium after overcrowding. T cells from aged mice, however, displayed significantly smaller levels of both resting and lectin-stimulated intracellular calcium concentration (p<0.01 each), as compared to those of the non-stressed, aged animals. This inadequate adaptation in the calcium metabolism of T lymphocytes may significantly contribute to the diminished immune response of the aged in stress.     

Changes in the T4/T3 molar ratio following thyrotropin releasing hormone injection in cattle.

The injection of thyrotropin releasing hormone into cattle resulted in a rapid decrease in the T4/T3 molar ratio. 2 breeds of cattle, Shorthorn and Africander Cross were studied. The decrease in the T4/T3 molar ratio was significantly greater in the Shorthorn breed. It is concluded that acute stimulation of the thyroid gland with TRH results in enhanced release of both T3 and T4 and that T3 is discharged more rapidly than T4.     

Distribution of intravenously administered acidic and basic fibroblast growth factors in the mouse.

Iodinated acidic or basic fibroblast growth factor (aFGF or bFGF) were separately injected into adult mice to follow their distribution in the main organs of the animals. Iodinated FGFs intravenously injected into mice cleared from blood with a T1/2 of 30 s. They mainly bound to kidney, liver and spleen. The binding of FGFs to these organs was maintained when the latter were washed with a physiological buffer containing 0.15 M NaCl, but it was eliminated when the buffer contained 2 M NaCl. Simultaneous injections of the FGFs together with increasing doses of heparin weakened the binding of FGF to vessels in a dose-dependent manner.     

Cartilaginous tissue and nalidixic acid]

1. Medication with nalidixic acid never produced articular lesions in adult animals. 2. In the species of immature animals, if sensitive to nalidixic acid, the articular lesions were macroscopically and microscopically identical. 3. The lesions regressed spontaneously as the basal germinative layer was never affected.     

Effect of a steroid contraceptive, "Enidrel" on the pre- and postnatal development of rats

To determine the long-term effects of prolonged and intensive treatment with an anticonceptive steroid, 60 female Wistar rats were given 1 mg/kg/day Enidrel (norethynodrel plus ethinyl estradiol 3-methyl-ether) by gastric sound for 60 days. The animals were divided into 2 groups and were placed with males for 10 days. Group 1 continued to recieve Enidrel through mating to the 15th day of gestation; Group 2 received no further treatment. General behavior and weight of the animals was unchanged when compared with controls. There were profound disturbances in the estrous cycle, but couplings and number of impregnations were normal in both groups. Most of the females in Group 1 aborted normal fetuses between Gestation Days 8 and 15; those in Groups 2 continued to term. Group 1 animals were remated and siblings of the 1st generation were crossed. The animals were fertile and the ne onates developed normally. It is concluded that Enidrel had no effect on morphogenesis and no masculinization effect on the hypophyso-hypothalamic system, as evidenced by the normal sexual behavior and fertility of the females.     

氧化型低密度脂蛋白诱导3T3-L1脂肪细胞内质网应激相关标志物p-eIF2a、CHOP表达

目的 观察氧化型低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导3T3-L1脂肪细胞内质网应激相关标志物p-eIF2a、CHOP的表达,探讨ox-LDL对3T3-L1脂肪细胞内质网应激的诱导作用.方法 体外培养3T3-L1脂肪细胞,用ox-LDL分别干预脂肪细胞6小时、12小时、24小时,用RT-PCR检测CHOP mRNA表达,用Western blot检测p-eIF2a、CHOP蛋白表达.结果 ox-LDL可诱导脂肪细胞p-eIF2a、CHOP表达,p-eIF2a表达发生在ox-LDL作用早期(6小时,12小时),24小时表达有所下降,而CHOP的表达强度随时间增加,24小时表达达到高峰.结论 ox-LDL可诱导3T3-L1脂肪细胞发生内质网应激,激活未折叠蛋白反应信号通路.p-eIF2a表达发生在ox-LDL作用早期,而CHOP的表达强度呈时间依赖性.     

Accelerated development of a Th-2 type factor in animals with and without an imbalance between help and suppression

Summary A helper factor can be detected in antigen-treated supernatants from spleen T and adherent cells of sensitized animals. This factor promotes an indirect hapten-specific plaque forming response of B cells, irrespective of the identity of the carrier, i.e. provides the Th-2 type of help. Factor production increases with age and occurs most rapidly in strains known to have an accelerated decrease of suppressor capacity. The reason for the inverse correlation between suppressor capacity and the Th-2 type of helper factor is discussed.Acknowledgments. Thanks are due to the Medical Research Council, the National Cancer Institute of Canada and the National Health Research and Development Program for financial support; T.M. is indebted to the Medical Research Council for personal support.     

Control of the hypothalamus-pituitary-adrenal axis in young and older rats injected with thyroxine.

Administration of T4 on alternate weeks for 30 weeks at a dosage which does not alter body weight depresses basal serum corticosterone levels in older rats (575 days), but not in young animals (260 days). Similar serum corticosterone response to HPA axis stimulation occurs regardless of age or T4 injection.     

Endogenous animal toxin-like human β-defensin 2 inhibits own K<Superscript>+</Superscript> channels through interaction with channel extracellular pore region

Human potassium channels are widely inhibited by peptide toxins from venomous animals. However, no human endogenous peptide inhibitor has been discovered so far. In this study, we demonstrate for the first time using electrophysiological techniques, that endogenous human β–defensin 2 (hBD2) is able to selectively and dose-dependently inhibit the human voltage-gated Kv1.3 channel at picomolar peptide concentration. The co-immunoprecipitation assays further supported the selective binding of hBD2 to Kv1.3 channel. Using mutagenesis experiments, we found that the outer pore domain of Kv1.3 channel was the binding site of hBD2, which is similar to the interacting site of Kv1.3 channel recognized by animal toxin inhibitors. The hBD2 was able to suppress IL-2 production through inhibition of Kv1.3 channel currents in human Jurkat cells, which was further confirmed by the lack of hBD2 activity on IL-2 production after Kv1.3 knockdown in these cells. More interestingly, hBD2 was also found to efficiently inhibit Kv1.3 channel currents and suppress IL-2 production in both human primary CD3⁺ T cells and peripheral mononuclear cells from either healthy donors or psoriasis patients. Our findings not only evidenced hBD2 as the first characterized endogenous peptide inhibitor of human potassium channels, but also paved a promising avenue to investigate newly discovered function of hBD2 as Kv1.3 channel inhibitor in the immune system and other fields.     

Organization in "barrels" of layer-IV cells of the S1 cortex in the mouse: effects of lesions or deprivation of the mystacial vibrissa]

In newborn mice the mystacial pads of the upper lips were submitted to different destructive or functionally reducing procedures. The effects on the barrels of the somato-sensory cortex were judged in mature animals and classified into 3 categories: 1. the barrels were present and normal; 2. the cortex was devoid of barrel structures; 3. the barrels were disorganized. Only procedures which involved peripheral lesions, and not those which reduced only the functional level, produced effects as deep as absence or disorganization of the barrels.     

Long-term 24-hour rest-activity pattern of sheep in stalls and in the field

Motor activity of sheep was continuously recorded for 2-3 weeks with an ambulatory monitoring device. Recordings were obtained from free-ranging animals in the field and from animals maintained under various controlled conditions in stalls. The sheep were diurnal under all conditions. While the daily amount of activity and the frequency of rest episodes showed only small differences between the conditions, the rest-activity pattern showed prominent differences. The pattern differed particularly between the field and the stalls. In the field, activity started to increase one hour after dawn, reaching a first maximum towards noon; a second, higher peak in the evening was followed by a rapid decline after dusk. In the stalls the onset and offset of activity was more abrupt; activity peaks coincided with feeding and human activity; the onset of rest with lights off. Activity was lowest and rest most prominent in those stalls where the animals were most isolated from human influence.     

Long-term 24-hour rest-activity pattern of sheep in stalls and in the field

Summary Motor activity of sheep was continuously recorded for 2–3 weeks with an ambulatory monitoring device. Recordings were obtained from free-ranging animals in the field and from animals maintained under various controlled conditions in stalls. The sheep were diurnal under all conditions. While the daily amount of activity and the frequency of rest episodes showed only small differences between the conditions, the rest-activity pattern showed prominent differences. The pattern differed particularly between the field and the stalls. In the field, activity started to increase one hour after dawn, reaching a first maximum towards noon; a second, higher peak in the evening was followed by a rapid decline after dusk. In the stalls the onset and offset of activity was more abrupt; activity peaks coincided with feeding and human activity; the onset of rest with lights off. Activity was lowest and rest most prominent in those stalls where the animals were most isolated from human influence.     

Enhancement of humoral immune response against human lung elastin peptides

Summary When guinea pigs instead of rabbits were used as the host animals, 8–16 times higher antibody titers against human lung elastin peptides were produced with only 1/20 the amount of antigen per unit body weight. This corresponds to a 200-fold enhancement of the immune response.Presented at the 6th Colloquium of the Federation of European Connective Tissue Clubs, Paris, August 28–30, 1978. The data form part of the Ph.D. thesis submitted by T.V. Darnule to the Department of Biology, New York University. Supported in part by NIH Program Project Grant HL15832 and by a Parker B. Francis Fellowship to T.V. Darnule.     

Experimental immunity against trypanosomiasis

Summary 9 groups of 6 female rats were used in an experiment using fraction 3 ofTrypanosoma rhodesiense. 500 g gave 100% immunoprotection and 1000 and 1500 g gave 66% immunoprotection when challenged with 5×10² T. brucei. 2 groups of 10 female rats were tested for a short period inoculation immune response. In this, 750 g of fraction 3 ofT. rhodesiense gave 70% immunoprotection when challenged withT. brucei.I thank the University of Zambia for support; Dr M. A. Q. Awan, Mazabuka, Zambia, forT. rhodesiense andT. brucei; Prof. J. W. Kibukamusoke, Dr O. Okong'o and Dr W. W. Anokbonggo for advice; Mr L. Nyaliti for technical help.     

Aufhebung der Thermothyrin-A-Sekretion der Schilddrüse durch Fütterung mit Methylthiouracil

Summary G. Mansfeld demonstrated that in the serum of overheated animals a substance (thermothyrine A) is present which, injected into normal animals, decreases O₂-consumption. Serum of thyroidectomized animals has no effect.Dogs and rabbits were treated daily with 0.10 g per kg methylthiouracil during 4 weeks, and were than subjected for 5 hours to a temperature of 34–35° C which raised their body temperature by 0.5–1.5° C. 2.5 cm³ of serum obtained at the end of the 5 hours period failed to reduce O₂-consumption of normal rats, while sera of untreated dogs and rabbits produced after similar exposure to high temperature a fall of O₂-consumption by 14–48%. It is therefore evident that methylthiouracil not only inhibits the formation of thyroxine but of thermothyrine A as well.The fact that thermothyrine A contains no iodine proves conclusively that the action of thiouracil compounds cannot be exclusively an inhibition of iodinization.     

Dexamethasone enhances CTLA-4 expression during T cell activation

T cell activation is enhanced by the costimulatory interaction of B7 on antigen-presenting cells and CD28 on T cells, resulting in long-term T cell proliferation, differentiation and production of large amounts of cytokines, such as interleukin (IL)-2. CTLA-4 is a co-stimulation receptor that shares 31% homology with CD28 and binds B7 family members with higher affinity. CTLA-4 is transiently expressed intracellularly and on the cell surface following activation of T cells. We have studied the kinetics of CTLA-4 expression and the effects of dexamethasone on CTLA-4 expression during T cell activation in cultures of mouse spleen cells stimulated by a mixture of immobilized anti-CD3 and anti-CD28 monoclonal antibodies (anti-CD3/CD28 mAb) or concanavalin A (ConA). CTLA-4 expression peaked on day 2 and returned to background levels after 7 days. Dexamethasone was found to potentiate CTLA-4 expression in a dose-dependent manner with an EC₅₀ effective concentration 50%) of about 10⁻⁸ M. In contrast, other immunosuppressive agents, such as rapamycin or cyclosporin A had no or an inhibitory effect on CTLA-4 expression, respectively. Dexamethasone also stimulated CD28 expression, but inhibited IL-2R expression during anti-CD3/CD28 mAb-induced mouse splenic T cell activation. Western blot analyses of lysates of activated mouse T cells showed that dexamethasone increased CTLA-4 protein levels twofold during anti-CD3/CD28 mAb-induced activation. Dexamethasone also enhanced CTLA-4 messenger RNA twofold as quantified by ribonuclease protection assay. The effects of dexamethasone on CTLA-4 expression were glucocorticoid-specific and completely inhibited by the glucocorticoid receptor antagonist mifepristone (RU486), indicating that the effect of dexamethasone on CTLA-4 expression is mediated through the glucocorticoid receptor. In conclusion, the immunosuppressive agent dexamethasone actually stimulates CTLA-4 expression, which is involved in downregulation of T cell activation. Received 19 May 1999; received after revision 13 July 1999; accepted 13 July 1999     

Serum-induced stimulation of snRNA synthesis in mouse 3T3 fibroblasts

Small nuclear RNAs (snRNAs) from quiescent and serum-stimulated 3T3 cultures, labeled with [3H]uridine [( 3H]U), were electrophoresed in polyacrylamide-urea slab gels and revealed by staining with ethidium bromide and by fluorography. Judged by labeling with [3H]U, synthesis of 7S and U1-U6 RNAs was very low or absent in quiescent cultures. The serum-induced transition of 3T3 cells from a resting to a growing state was accompanied by an early, apparently sequential stimulation of snRNA synthesis; stimulated synthesis of 7S, U1, U2, U3, U4 and U6 RNAs coincided in time with serum-induced stimulation of 45S pre-ribosomal RNA (pre-rRNA) and heterogeneous nuclear RNA (hnRNA) synthesis.