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.     

Absorption of iron from gut into blood: Sex- and time-related studies in rats

Summary As judged from 2-h blood level curves, adult female rats absorbed more Fe^II per cm² of gross duodenal mucosa than adult male rats. By contrast, the 2-h blood level curves per cm² of mucosa of proximal jejunum did not differ significantly in male and female rats although in both sexes, iron was absorbed more efficiently from the duodenum. This research was supported by grant RO1 AM 12381 from the National Institute of arthritis, Diabetes and Digestive and Kidney Diseases, National Institutes of Health.     

Effect of temperature on in vitro proliferative activity of human umbilical vein endothelial cells

Human umbilical vein endothelial cells, skin fibroblasts, and retinal pigment epithelial cells are cultivated in medium supplemented with 15 to 20% serum in our laboratory. The effects of various incubation temperatures on the proliferation of these cells was examined. Our study shows that the mitogenic response of the endothelial cells to a change of temperature differed markedly from that of the fibroblasts and epithelial cells. Cultivation of human umbilical vein endothelial at 37°C required seeding densities as high as 1–2×10⁴ cells/cm², and yet resulted in a low growth rate and premature senescence. However, under the same culture conditions, but at 33°C, the proliferative capacity of these endothelial cells was potentiated. The results were striking; at 33°C the cells grew actively and the life span was extended. The number of cumulative population doublings increased fourfold compared with that for the same cells cultivated at 37°C. The inoculum size could be reduced, since at 33°C the endothelial cells were able to replicate at seeding densities as low as 20 cells/cm². The cells serially subcultured at 33°C retained morphological features and specific immunological markers of endothelial cells.     

The influence of cold or isolation stress on resistance of mice to West Nile virus encephalitis

Summary The effect of cold or isolation stress on mortality rate and brain virus level were investigated in mice infected with West Nile virus (WNV). Exposure of mice for 5 min/day to cold water (1±0.5°C) for 8–10 days resulted in 92% mortality as compared to 47% in control mice (p<0.001). Mice housed in individual cages (isolation stress) were also more susceptible to WN viral infection, as shown by increased mortality rate reaching 85% as compared to 50% in mice housed 6 per cage (p<0.01). Cold or isolation stress increased blood brain and spleen virus levels as early as 2 days after inoculation. After 8 days of isolation or cold stress, mice inoculated with WNV had 8.9 and 9.0 log₁₀ plaque forming units in the brain, respectively, as compared to 6.9 in the control (p<0.01–0.001). Furthermore, lymphoid organs such as spleen and thymus showed severe mass loss. These data suggest that physical or non-physical stress situations enhance WNV encephalitis by accelerating virus proliferation and increase mortality in mice.     

An H-2-associated difference in murine serum cholesterol levels

Summary We describe, in mice, a difference in serum cholesterol and adrenal weight associated with an H-2^a/H-2^b haplotype difference.     

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.     

Fehlerkorrekturen bei stereologischen Messungen an menschlichen Plazenten: 1. Die Schrumpfung

Summary In sterological measurings on human placental chorionic villi the measured volume per cm³ is shown not to be influenced by shrinking of the tissue. The measured villous surface per cm³, however, must be corrected by multiplication with the factor 1–p, wherep means the percentage of linear shrinking expressed as decimal fraction.

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Arbeit mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Drug susceptibility of PCA in WBB6F1-W/Wv mice

Our previous study revealed that passive cutaneous anaphylaxis (PCA) can be produced in congenitally mast cell-deficient WBB6F1-W/W^v (abbreviated as W/W^v) mice on sensitization with undiluted or slightly diluted allogeneic and xenogeneic antisera but not on sensitization with allogeneic monoclonal immunoglobulin (Ig)E and IgG1 antibodies regardless of the antibody concentration [1]. In view of these findings, the present study was conducted to characterize PCA in this strain from its drug susceptibilities using mast cell-bearing WBB6F1-+/+ (abbreviated as +/+) and B6D2F1 mice as references. PCA in W/W^v mice mediated by a low dilution (1  4) of hyperimmune serum to bovine serum albumin of the B6D2F1 mouse origin was markedly suppressed by CV-6209, an antagonist of platelet-activating factor (PAF), but not by antihistamines such as cyproheptadine and oxatomide. In contrast, PCA in +/+ and B6D2F1 mice mediated by a high dilution (1  128) of the anti-serum (virtually by IgG1 antibody) was nearly completely suppressed by antihistamines but not by CV-6209. A remarkable difference between PCA in W/W^v and reference mice was also observed in the susceptibility to monoclonal anti mouse granulocyte (Gr-1) antibody PCA in W/W^v mice was potently suppressed by the 1- to 3-day pretreatment with this antibody but that in references was not at all. Putting these present results together with the previous finding that anti-granulocyte antibody greatly reduces circulatory Gr-1⁺ leukocytes, 1 to 3 days after the treatment [2], it is highly probable that PCA in W/W^v mice mediated by some antibody isotypes other than IgE and IgG1 is produced by PAF mainly released from Gr-1⁺ cells, while IgG1 antibody-mediated PCA in mast cell-bearing reference mice is evoked by histamine derived from mast cells. PCA homologous to that in W/W^v mice could also be produced in the reference mice on sensitization with undiluted or slightly diluted antiserum, when generalized blueing due to excess IgG1 antibody was removed by the oxatomide treatment be fore the antigen challenge. Received 10 December 1997; received after revision 2 February 1998; accepted 23 February 1998     

Untersuchungen zur biologischen Wirksamkeit eines A-homo-Testosteronacetates

Summary The biological efficiency of a high purified A-homo-testosterone-acetate was tested on male mice. We found no inhibition of the gonadotropic partial-function of the anterior pituitary by the substance; and furthermore no substitution of testosterone deficit after castration was possible.     

Hormonal induction of steroid sulphatase in the mouse

Summary By comparing steroid sulphatase levels per se, and also ratios to -galactosidase, in 6 sets of mice — normal females, entire and castrated males both with and without exogenous testosterone administration — we obtained support for the contention that induction of this enzyme is in part controlled by male hormones.     

Pharmacological inhibition of store-operated calcium entry in MDA-MB-468 basal A breast cancer cells: consequences on calcium signalling,cell migration and proliferation

Store-operated Ca²⁺ entry is a pathway that is remodelled in a variety of cancers, and altered expression of the components of store-operated Ca²⁺ entry is a feature of breast cancer cells of the basal molecular subtype. Studies of store-operated Ca²⁺ entry in breast cancer cells have used non-specific pharmacological inhibitors, complete depletion of intracellular Ca²⁺ stores and have mostly focused on MDA-MB-231 cells (a basal B breast cancer cell line). These studies compared the effects of the selective store-operated Ca²⁺ entry inhibitors Synta66 and YM58483 (also known as BTP2) on global cytosolic free Ca²⁺ ([Ca²⁺]_CYT) changes induced by physiological stimuli in a different breast cancer basal cell line model, MDA-MB-468. The effects of these agents on proliferation as well as serum and epidermal growth factor (EGF) induced migration were also assessed. Activation with the purinergic receptor activator adenosine triphosphate, produced a sustained increase in [Ca²⁺]_CYT that was entirely dependent on store-operated Ca²⁺ entry. The protease activated receptor 2 activator, trypsin, and EGF also produced Ca²⁺ influx that was sensitive to both Synta66 and YM58483. Serum-activated migration of MDA-MB-468 breast cancer cells was sensitive to both store-operated Ca²⁺ inhibitors. However, proliferation and EGF-activated migration was differentially affected by Synta66 and YM58483. These studies highlight the need to define the exact mechanisms of action of different store-operated calcium entry inhibitors and the impact of such differences in the control of tumour progression pathways.     

Actionin vitro de la pénicilline sur la virulence du Tréponème pâle

Summary The Ghent strain ofTreponema pallidum loses entirely its virulence for the rabbit when mixed invitro during 1 hour at 18° C or 3 hours at 37° C with a solution of penicillin sodium Chas. Pfizer & Co., Inc. or with penicillin C.S.C. sodium salt of the Commercial Solvent Corporation at the rate of 10,000 Oxford units per cm³. The injection of 20,000 units in each testicle of the animal, when made immediately before or at the same time as the inoculation of the virus, does not protect against the infection. It is therefore possible to prevent syphilis by penicillizing the blood for transfusion. But too great amounts of the drug are required in practice. It must also be proved that chemically pure penicillin has the same treponemicid action.     

Cytosolic calcium in platelet activation

Experiments with permeabilised platelets, and with intact platelets loaded with fluorescent Ca²⁺-indicators, over the past several years have greatly extended our knowledge and understanding of cytosolic Ca²⁺ as a platelet activator and its interactions with other cytosolic regulators. This article outlines insights, gained from the use of the fluorescent dyes, into maintenance and restoration of basal [Ca²⁺]_i, mechanisms of receptor-mediated Ca²⁺-mobilisation and quantitation of [Ca²⁺]_i/response relations in intact human platelets.     

Serine protease PRSS55 is crucial for male mouse fertility via affecting sperm migration and sperm–egg binding

Testis-specific PRSS55 is a highly conserved chymotrypsin-like serine protease among mammalian species. So far, the physiological function of PRSS55 remains unknown. Here, we show that PRSS55 is a GPI-anchored membrane protein, specifically expressed in adult mouse testis and mainly observed in the luminal side of seminiferous tubules and sperm acrosome. Mice deficient for Prss55 develop male infertile with normal reproduction-related parameters observed. Interestingly, in vivo fertilization rate of Prss55^?/? males is dramatically decreased, possibly due to incapable migration of Prss55^?/? sperm from uterus into oviduct. However, in vitro fertilization rate has no difference between two genotypes although Prss55^?/? sperm presents defective recognition/binding to zona-intact or zona-free oocytes. Further study reveals that mature ADAM3 is almost undetectable in Prss55^?/? sperm, while precursor ADAM3 remains unchanged in the testis. However, it is shown that ADAM3 has no interaction with PRSS55 by immunoprecipitation with anti-PRSS55 antibody. The expression levels of several proteins known to be related to the observed phenotypes remain comparable between wt and Prss55^?/? mice. Moreover, we found that Prss55 deficiency has no effect on PRSS37 or vice versa albeit two mutant males share almost the same phenotypes. Microarray analysis reveals a total of 72 differentially expressed genes in Prss55^?/? testis, most of which are associated with cellular membrane and organelle organization, protein transport and complex assembly, and response to stimulus and signaling. In conclusion, we have demonstrated that PRSS55 plays vital roles in regulating male fertility of mice, including in vivo sperm migration and in vitro sperm–egg interaction, possibly by affecting the maturation of ADAM3 in sperm and the expression of multiple genes in testis.     

Beta-carotene affects gene expression in lungs of male and female <Emphasis Type="Italic">Bcmo1</Emphasis><Superscript>−/−</Superscript> mice in opposite directions

Molecular mechanisms triggered by high dietary beta-carotene (BC) intake in lung are largely unknown. We performed microarray gene expression analysis on lung tissue of BC supplemented beta-carotene 15,15′-monooxygenase 1 knockout (Bcmo1 ^−/−) mice, which are—like humans—able to accumulate BC. Our main observation was that the genes were regulated in an opposite direction in male and female Bcmo1 ^−/− mice by BC. The steroid biosynthetic pathway was overrepresented in BC-supplemented male Bcmo1 ^−/− mice. Testosterone levels were higher after BC supplementation only in Bcmo1 ^−/− mice, which had, unlike wild-type (Bcmo1 ^+/+) mice, large variations. We hypothesize that BC possibly affects hormone synthesis or metabolism. Since sex hormones influence lung cancer risk, these data might contribute to an explanation for the previously found increased lung cancer risk after BC supplementation (ATBC and CARET studies). Moreover, effects of BC may depend on the presence of frequent human BCMO1 polymorphisms, since these effects were not found in wild-type mice.     

Protective effect of Shigyaku-to,a traditional Chinese herbal medicine,on the infection of herpes simplex virus type 1 (HSV-1) in mice

The antiviral activity of Shigyaku-to (TJS-109), a traditional Chinese herbal medicine, was investigated in mice infected with herpes simplex virus type 1 (HSV-1). TJS-109 is a combination of the medicinal plant extracts fromZingiberis siccatum rhizoma,Aconiti tuber andGlycyrrhizae radix in a specific proportion. Mice infected with a 10 LD₅₀ dose of HSV-1 were treated with TJS-109 orally at doses of 1.25 to 20 mg/kg 2 days before, and 1 and 4 days after the infection. The treated groups had 80% (1.25 mg/kg), 40% (5 mg/kg) and 23% (20 mg/kg) mortality rates 25 days after the infection as compared with a 100% mortality rate in control mice treated with saline. When HSV-1 infected mice (recipients) received CD8⁺T cell fractions derived from spleens of mice treated with TJS-109 (donors), 70% of recipients survived, as compared with 0% survivors in the groups of mice treated with saline, B cell fractions, CD4⁺ T cell fractions or macrophage-enriched fractions prepared from the same donors. TJS-109 did not show any virucidal activities against HSV-1 or any virostatic activities on the growth of HSV-1 in Vero cells. These results suggest that TJS-109 protected mice exposed to lethal amounts of HSV-1 through the activation of CD8⁺ T cells.     

D-半乳糖致衰老小鼠学习记忆能力和睾丸的改变及延衰合剂的治疗作用

目的研究D-半乳糖所致亚急性衰老小鼠学习记忆能力和睾丸的变化,并观察延衰合剂对其的治疗作用。方法选昆明系雄性小鼠,用D-半乳糖建立亚急性衰老模型。应用Morris水迷宫实验测试衰老小鼠学习记忆能力的变化;电镜技术观察延衰合剂治疗后衰老小鼠睾丸的形态学改变;酶联免疫分析法检测血清睾酮的变化。结果衰老小鼠学习记忆能力下降,睾丸重量减少,生精细胞减少,结构紊乱,功能障碍,血清睾酮含量明显降低。在改变学习记忆能力上延衰合剂高剂量组作用明显优于延衰合剂低剂量纽（P〈0．05）,但与补肾益寿胶囊纽及延衰舍剂中荆量组差畀无显著性（P〉0．05）。结论延衰舍剂可以提高衰老小鼠学习记忆能力,改善睾丸生精小管的超微结构,抑制衰老小鼠血清睾酮含量的下降,具有一定的延缓衰老作用。     

Membrane constants and sodium conductance of a single muscle fibre

Riassunto Nella singola fibra muscolare di cavia a 38°C sono state determinate, applicando la «square pulse analysis», la resistenza, la costante di tempo, la costante di spazio e la capacità della membrana a riposo. È stato inoltre possibile calcolare la massima conduttanza per il sodio,gn a, che è risultata di 35,3 mmho/cm².     

Transgenic animal models of sickle cell disease

An animal model which allows study of chronic processes occurring in sickle cell disease has finally been realized with the development of several lines of transgenic mice which express high levels of ^s or ^s-variants in their red cells. The red cells of all mouse lines exhibit characteristic sickle shapes on deoxygenation and most lines have enlarged spleens and mildly elevated reticulocytes in adult mice; both of these properties are suggestive of enhanced red cell destruction and erythropoiesis. However, all lines examined to date have normal hemoglobin levels in adult mice. In one mouse line under ambient conditions, retinopathy and abnormal renal function have been observed and in the same line under hypoxic conditions, anemia, irreversibly sickled cell formation, and urine concentrating defect have been observed. The current mouse lines will allow some aspects of sickle cell disease to be studied, but sigificant improvements can still be made in the transgenic mouse model.     

Conversion of testosterone to androstenedione by liver homogenates of testicular feminized mice

Summary Liver homogenates of testicular feminized (Tfm) mice carrying the protective (o^hv) gene were found to be less capable of converting testosterone to androstenedione than Tfm without the protective gene.This work was supported by the Medical Research Council of Canada, No. MT 4192 and Ortho Pharmaceutical Company.