Cell-to-cell transfer of interferon-induced antiproliferative activity

Interferon-treated cells rapidly and efficiently transferred the antiproliferative activity of interferon to untreated cells. This phenomenon was not due to the carry-over of interferon by the interferon-treated cells. Thus, to evoke an antiproliferative state, interferon did not directly contact each cell in a population. The results suggest a novel mechanism by which interferon may indirectly regulate cell growth, and suggests that cells other than those of the immune system may play a role in controlling tumor growth in tissue where cell-to-cell contact occurs.     

Strain dependence of the antiproliferative action of interferon on murine erythroid precursors

Electrophoretically pure mouse interferon inhibits erythropoietin-dependent proliferation of committed erythroid precursors (CFU-E) obtained either from adult mouse bone marrow or from 14-day fetal mouse livers. The degree of inhibition is significantly influenced by the genotype of the cell donor; about ten times as much interferon is required to inhibit proliferation of CFU-E from C57BL/6 than is needed for comparable inhibition of CFU-E from BALB/c or Swiss mice. These strain-dependent results point to the existence of genes that influence the degree of the inhibitory effect of interferon on cell multiplication.     

Inhibition of development of exoerythrocytic forms of malaria parasites by gamma-interferon

A specific DNA probe was used to study the effect of recombinant rat, mouse, and human gamma-interferon (gamma-IFN) on the course of sporozoite-induced malaria infections. In mice and rats infected with sporozoites of Plasmodium berghei, mouse and rat gamma-IFN's strongly inhibited the development of the exoerythrocytic forms in the liver liver cells of the hosts, but not the development of the erythrocytic stages. The degree of inhibition of the exoerythrocytic forms was proportional to the dose of gamma-IFN administered, but was independent of the number of sporozoites used for challenge. A 30 percent reduction in the development of exoerythrocytic forms in rat liver was achieved when 150 units (about 15 nanograms of protein) of rat gamma-IFN were injected a few hours before sporozoite challenge; the reduction was 90 percent or more with higher doses of gamma-IFN. The effect was less pronounced if the gamma-IFN was administered 18 hours before or a few hours after challenge. Human gamma-IFN also diminished the parasitemia in chimpanzees infected with sporozoites of the human malaria parasite Plasmodium vivax. The target of gamma-IFN activity may be the infected hepatocytes themselves, as shown by in vitro experiments in which small doses of the human lymphokine inhibited the development of exoerythrocytic forms of Plasmodium berghei in a human hepatoma cell line. These results suggest that immunologically induced interferon may be involved in controlling malaria infection under natural conditions.     

Nicotinic antagonists enhance process outgrowth by rat retinal ganglion cells in culture

Functional nicotinic cholinergic receptors are found on mammalian retinal ganglion cell neurons in culture. The neurotransmitter acetylcholine (ACh) can be detected in the medium of many of these retinal cultures, after release presumably from the choline acetyltransferase-positive amacrine cells. The postsynaptic effect of endogenous or applied ACh on the ganglion cells can be blocked with specific nicotinic antagonists. Here it is shown that within 24 hours of producing such a pharmacologic blockade, the retinal ganglion cells begin to sprout or regenerate neuronal processes. Thus, the growth-enhancing effect of nicotinic antagonists may be due to the removal of inhibition to growth by tonic levels of ACh present in the culture medium. Since there is a spontaneous leak of ACh in the intact retina, the effects of nicotinic cholinergic drugs on process outgrowth in culture may reflect a normal control mechanism for growth or regeneration of retinal ganglion cell processes that is exerted by ACh in vivo.     

Interferon: production by chick erythrocytes activated by cell fusion

In the presence of Sendai virus inactivated with ultraviolet light, nucleated chick erythrocytes can be fused with several types of human cells to form heterokaryons. Although chick erythrocytes alone cannot be stimulated by Sendai virus to produce interferon, fusion with a human cell (AH-1) which itself may produce human interferon results in heterokaryons in which the erythrocyte genome is activated and chick interferon is produced. When nucleated chick erythrocytes are fused with another type of human cell (HeLa clone S-3) which does not produce human interferon when stimulated, no chick interferon is detectable, despite morphologic changes suggestive of activation of the erythrocyte nuclei.     

Interferon binding: the first step in establishment of antiviral activity

Chick cells incubated at 1 degrees C with interferon fail to develop antiviral activity, but this activity appears subsequent to a 7-hour incubation at 37 degrees C after removal of interferon by repeated washings. Treatment with actinomycin D blocks the development of the latter activity. Cells incubated with interferon at 1 degrees C for up to 1 hour and then washed and incubated for 2 hours at 37 degrees C develop a degree of antiviral activity proportional to the concentration of interferon at initial incubation; at any concentration, the antiviral activity increased with the duration of initial incubation at 1 degrees C, but a maximal response was reached at 10 or 20 minutes. Treatment with trypsin after incubation with interferon at 1 degrees C inhibited development of antiviral activity. Interferon is rapidly bound to a superficial cell site, and this binding is necessary for development of antiviral activity in chick cells.     

动物细胞凋亡与疾病的发生

细胞凋亡和细胞坏死是细胞死亡的两种形式，前者属于生理性的平衡机制，后者则是由于致病因子作用下发生的病理过程。免疫系统抑制与细胞凋亡有关，在免疫系统中存在两种细胞程序性死亡，一种是在无生长因子时，Bcl-2基因调控的程序化死亡；另一种是细胞毒介导的靶细胞的程序化死亡。细胞凋亡是动物机体对病毒侵入的一种防御机制之一，当细胞凋亡和细胞增殖平衡破坏时，可引起多种疾病的发生。     

Chloramphenicol: effects on mouse myeloma cells in tissue culture

Within 36 hours of being administered, chloramphenicol (50 micrograms per milliliter) inhibits by 50 percent the rate of protein synthesis in mouse myeloma cells grown in suspension culture. Although there is a decrease in the amount of globulin synthesized, the rate of synthesis per cell is unchanged; the observed decrease is traced to the inhibition of cell proliferation caused by chloramphenicol.     

大蒜对铜绿微囊藻的化感抑制作用

通过人工培养箱培养的方法,利用分光光度计、酶标仪及相关试剂盒等分别测定铜绿微囊藻在不同浓度(0,0.2,0.4,0.8,1.6,3.2,6.4 g/L)大蒜作用下的生理生化指标变化情况,以期探讨大蒜对铜绿微囊藻的抑制机理。结果表明:大蒜对铜绿微囊藻表现为低促高抑的作用效果,与对照相比,0.2 g/L的大蒜浓度对铜绿微囊藻生长反而略有促进作用;6.4 g/L浓度的大蒜则可有效地抑制铜绿微囊藻的生长,而且随着作用时间的延长其抑制效果愈加明显,在第96小时时铜绿微囊藻的生长抑制率可达53.42%;另外,高浓度大蒜能有效抑制铜绿微囊藻的叶绿素a含量并能影响铜绿微囊藻藻胆蛋白各组分的构成及可溶性蛋白的含量。由此可以得出,一定浓度的大蒜对铜绿微囊藻具有化感抑制作用,且高浓度下的大蒜能通过影响铜绿微囊藻叶绿素a的含量来影响其光合作用,最终导致藻类繁殖受阻。可见,藻类叶绿素a(即PSⅡ)是大蒜抑制铜绿微囊藻生长繁殖的一个作用位点。     

鸡重组α干扰素与鸡重组γ干扰素抗新城疫病毒活性研究

将鸡IFN-α和IFN-γ原核表达产物经Ni2＋-NTA亲和层析法纯化,进行SDS-PAGE分析,再将纯化并已测定浓度的鸡重组IFN-α和IFN-γ稀释到1/10倍;用细胞病变抑制法检测重组表达蛋白的抗新城疫病毒活性,并利用感染病毒的鸡胚来测定鸡重组表达蛋白的抗病毒活性。结果显示,SDS-PAGE可检测到相对分子质量为25kU左右的蛋白条带,纯化的鸡IFN-α和IFN-γ抗新城疫病毒复制的活性分别为4.32×103 U/mg和2.65×103 U/mg;当鸡IFN-α和IFN-γ的比例为125∶1时,其联合抗新城疫病毒复制的活性最佳。抗鸡胚感染病毒活性的测定结果表明,与病毒对照组相比,IFN-α和IFN-γ及IFN-α和γ联合组,HA滴度都有明显的降低;并且从鸡胚的平均死亡时间上,联合干扰素对新城疫病毒的抑制作用更强于单个干扰素的应用。这证明了鸡IFN-α、IFN-γ及其联合干扰素对新城疫病毒的复制有明显的抑制作用。     

Interferon production and action in mouse, hamster and somatic hybrid mouse-hamster cells

A hybrid mouse-hamster cell line was developed from a mouse cell line which produces a high titer of interferon and is sensitive to its action, and a hamster cell line which produces little interferon and is relatively insensitive to its action. Parental cell lines demonstrated complete species specificity with respect to interferon production and action. The hybrid cells produced interferon (or interferons) effective when tested on the mouse cell line and primary hamster cells; the hybrids were sensitive to the action of both mouse and hamster interferons. Hybrid cells produced ten times more hamster interferon than the parent hamster cell line and were eight times more sensitive to hamster interferon than the parent hamster cell line.     

Adenovirus tumorigenesis: role of the viral genome in determining tumor morphology

Adenovirus type 12 transforms the fibroblastic BHK21 (baby hamster kidney) cell line into rounded or cuboidal cells that give rise in hamsters to undifferentiated small cell sarcomas indistinguishable from those induced in newborn hamsters by inoculation of the virus itself. In contrast. cells from this line transformed by polyoma virus retain their fibroblastic morphology and induce fibrosarcomas in hamsters. This suggests that the morphology of tumors induced by the adenovirus-transformed cells from this line may be determined by the viral genome and that such mechanism may also explain the remarkably uniform microscopic appearance which seems to characterize tumors induced in hamsters by direct inoculation of adenovirus type 12.     

Viral infection and host defense

Double-stranded RNA, made as an intermediary substance in the replication of most, if not all, viruses, may play a much more important role in the pathogenesis and the recovery from virus infections than has hitherto been suspected. Apparently, dsRNA is used by both the challenge virus and the host cell in an attempt to gain "molecular control." Double-stranded RNA exerts a set of effects, which may be well balanced, not only at the level of the individual cell but also at the complex assemblage of these cells termed the organism (Fig. 1). In the cell, interferon synthesis is triggered, although interferon mRNA translation may not occur if dsRNA shuts off protein synthesis too quickly. In the whole organism, the disease severity will depend on how certain toxic reactions evoked by infection (such as cell necrosis and fever) are counterbalanced by an increase in the host defense mechanisms (for example, immune responsiveness and interferon production). Many aspects of the response, relating to either progress of, or recovery from, the disease, can be explained on the basis of a dsRNA. In addition to drawing attention to the biodynamic role of dsRNA, our hypothesis suggests specific experimental vectors designed to enhance our information on the molecular basis of the morbid process which occurs with viral infection. Finally, we suggest that, although the dsRNA molecule may be viewed as a rather simple unit structure, the opportunity for further diversity in the biological activity of a given dsRNA molecule always exists. Namely, each deviation from a perfectly double-helical arrangement introduces the possibility for emphasizing one biological reactivity at the expense of another. This latter structure-activity property may partially account for the extreme apparent diversity, commonly encountered, in the presentations of virologic illness. Appendix note added in proof. Subsequent to submission of this text, we have found that the potent mitogen effect of dsRNA for lymphocytes (murine and human) is also exquisitively sensitive to the fidelity in base pairing of the input polymer pair (59). For example, infrequent "loops" (one nucleotide per 20 base pairs) in an otherwise perfectly helical rI(n) (.) rC(n) molecule [for example, rI(n) (.) r(C(19,)U)(n)] strongly changes its mitogenic properties. This observation, which supports our thesis that a "fine structure" term can be developed for other reactions triggered by dsRNA's in biological systems, emphasizes that diverse biological effects may be encountered with an ostensibly uniform family of dsRNA's.     

Inhibition of endothelial regeneration by type-beta transforming growth factor from platelets

Damage to the vessel wall is a signal for endothelial migration and replication and for platelet release at the site of injury. Addition of transforming growth factor-beta (TGF-beta) purified from platelets to growing aortic endothelial cells inhibited [3H]thymidine incorporation in a concentration-dependent manner. A transient inhibition of DNA synthesis was also observed in response to wounding; cell migration and replication are inhibited during the first 24 hours after wounding. By 48 hours after wounding both TGF-beta-treated and -untreated cultures showed similar responses. Flow microfluorimetric analysis of cell cycle distribution indicated that after 24 hours of exposure to TGF-beta the cells were blocked from entering S phase, and the fraction of cells in G1 was increased. The inhibition of the initiation of regeneration by TGF-beta could allow time for recruitment of smooth muscle cells into the site of injury by other platelet components.     

大豆异黄酮对金黄色葡萄球菌的抑菌机制研究

 【目的】研究大豆异黄酮（Soybean isoflavone,SI）的抑菌机制。【方法】本试验利用呼吸代谢抑制实验和4',6-二脒基-2-苯基吲哚（4',6-diamidino-2-phenylindole,DAPI）荧光染色对大豆异黄酮的抑菌机制进行研究。【结果】呼吸代谢抑制实验结果表明,当SI的终浓度为0.8 mg?ml-1时,可显著抑制金黄色葡萄球菌苹果酸脱氢酶的活性,SI组苹果酸脱氢酶的比活性与对照组相比降低了74.97%。DAPI染色结果显示,DAPI可以和金黄色葡萄球菌内的核酸结合,经过SI作用28 h后,菌体内的核酸含量呈明显下降趋势,其中DNA的荧光强度减少了33.53%,RNA减少了39.82%。【结论】SI具有有较强的抑菌活性,其抑菌机制是通过抑制细菌的呼吸代谢和核酸的合成等方面实现的。     

CIRCULATING INTERFERON IN MICE AFTER INTRAVENOUS INJECTION OF VIRUS

Circulating interferon was detectable in mouse serum within 1 hour after the intravenous injection of various types of virus and it reached maximum levels in about 4 hours. Rapidly produced interferon may play a role in the pathogenesis of viral infection and in viral interference.     

Fish oils inhibit endothelial cell production of platelet-derived growth factor-like protein

Diets rich in fish and fish oils are associated with a reduced risk of cardiovascular disease and atherosclerosis. The interaction of a commercial fish oil extract (MaxEPA) with vascular endothelial cells (ECs) was studied as a possible mechanism for this protective effect. MaxEPA almost completely inhibited EC production of platelet-derived growth factor-like protein (PDGFc) while other lipids had a lesser effect or no effect. Overall protein synthesis was not reduced, nor was the inhibition due to defective secretion or increased degradation of the growth factor. Antioxidants suppressed the inhibitory activity of MaxEPA indicating that free radical oxidative processes were required for the inhibition. These results suggest that fish oils may suppress intimal smooth muscle cell proliferation by decreasing the production of EC-derived paracrine growth factors. This inhibitory process represents a possible molecular mechanism for the antiatherosclerotic action of marine lipids.     

Nuclear-cytoplasmic interaction in DNA synthesis

In Amoeba proteus the transplantation of a nucleus engaged in DNA synthesis into a G(2)-phase (after DNA synthesis) cell results in inhibition of such synthesis. When the nucleus of a G(2) cell is transplanted into an S-phase (period of DNA synthesis) cell, such a nucleus may begin to synthesize DNA.