The use of 5-fluorocytosine and ketoconazole in the culture of the erythrocytic stages of Plasmodium falciparum and some tumor cell lines

In vitro culture systems are often contaminated by bacteria and fungi. It is therefore often necessary to supplement culture media with agents such as penicillin/streptomycin, gentamycin or amphotericin B. The latter cannot be used in the in vitro culture of erythrocytic stages of P. falciparum, and thus anti-fungal agents have not been regularly used in this system. We describe the prophylactic use of 5-fluorocytosine (5-FC) and ketoconazole (KTZ) in tissue cultures at concentrations up to 300 and 10 micrograms/ml respectively which have no effect on the growth of P. falciparum (FCR-3 strain). A melanoma cell line (C32) and a line of uterine carcinoma (C41) were also unaffected by similar concentrations of 5-FC and KTZ. When dissolved in complete culture medium (RPMI 1640) with 10% human plasma, the minimum inhibitory concentration of 5-FC for a susceptible strain of Candida remained below 2 micrograms/ml. These experiments suggest that 5-FC (at 50 micrograms/ml) alone or in combination with KTZ (at 1 microgram/ml) is a useful addition to the armamentarium of antimicrobials available to the tissue culture biologist for a variety of cell culture systems.     

Phenytoin-induced DNA synthesis and inositol 1,4,5-trisphosphate formation in L-929 fibroblasts

Culture of L-929 fibroblasts in the presence of phenytoin (2.5-5.0 micrograms/ml) increased DNA synthesis, as indicated by increased [3H]thymidine uptake, while a higher dose (20 micrograms/ml) inhibited DNA synthesis. In like manner, a low dose of phenytoin (5.0 micrograms/ml) was effective in increasing inositol 1,4,5-trisphosphate formation while a higher dose (10 micrograms/ml) tended to inhibit this activity. These data suggest that the formation of inositol phosphate second messengers may play a role in phenytoin-induced fibroblast proliferation and connective tissue growth.     

Isometachromin, a new cytotoxic sesquiterpenoid from a deep water sponge of the family Spongiidae.

Isometachromin (1), a new sesquiterpene-quinone that is related structurally to metachromin C (2), and the known compounds ilimaquinone (3) and 5-epi-ilimaquinone (4), were isolated from a deep water sponge in the family Spongiidae; the structure of isometachromin was elucidated by spectral methods. Isometachromin exhibits in vitro cytotoxicity against the human lung cancer cell line A549 (IC50 = 2.6 micrograms/ml), but not against P388 murine leukemia (IC 50 > or equal to 10 micrograms/ml) and also exhibits antimicrobial activity.     

Failure of somatostatin to influence experimental tumor cell growth in vivo and in vitro

The influence of somatostatin on tumor cell growth was studied in vivo in mice (sarcoma 180 ascites tumor and Lewis lung tumor) and in vitro on nontransformed and polyoma-transformed cell lines. 4 or 20 micrograms/100 g of cyclic somatostatin and 4 micrograms/100 g of linear protamin Zn-bound somatostatin were injected s.c. twice daily in the in vivo study. Cyclic somatostatin (1, 4 or 10 micrograms/ml) was added twice daily to the cell cultures. Somatostatin administration influenced neither the survival of animals nor the growth rate of cultured cell lines.     

Persistence of added retinoids in organ culture media during induction of mucous metaplasia and glandular morphogenesis in hamster cheek pouches

The retinoid concentration (determined colorimetrically) did not change significantly in retinyl acetate-supplemented (6 micrograms/ml) Eagle's Minimal Essential Medium containing 10% fetal calf serum when stored at -20 or 4 degrees C over 7 days. After the medium was incubated at 37 degrees C for 48 h, 37-49% of the retinoid remained, whether or not tissue (neonatal Syrian hamster cheek pouch) was present, and irrespective of explant age. The normal retinoid level in the tissue was approximately 0.25 micrograms per gram. Therefore, neonatal hamster cheek pouches, incubated in medium with the addition of 6 micrograms of retinyl acetate per ml of medium and undergoing mucous metaplasia and some mucous gland morphogenesis, were continually being exposed to retinoid levels which, though gradually decreasing, remained well above their normal physiological level.     

Exogenous fibronectin modifies the aggregation of collagen-stimulated human platelets

Fibronectins (FN) are adhesive glycoproteins whose role in platelet aggregation is unclear. Addition of 3, 6 and 12 micrograms/ml of human plasma FN in vitro to isolated human platelets, which had been freed from plasma FN by gel filtration and subsequently stimulated with collagen, inhibited the last stage of platelet aggregation. With 3 and 6 micrograms/ml of FN a shortening of the lag-time was also observed. These data showed that FN may play a role in platelet-collagen interaction as well as in platelet-platelet interaction.     

The effect of nicotine on ethanol-induced gastric ulcers in rats

Nicotine, in concentrations of 5 and 25 micrograms/ml drinking water, given ad libitum for 10 days, dose-dependently increased lesion formation and worsened ethanol-induced ulceration in rat stomachs. Daily fluid intake and b.wt gain were not adversely affected by nicotine pretreatment.     

Elevated glucose levels influence in vitro hatching,attachment, trophoblast outgrowth and differentiation of the mouse blastocyst

Summary A glucose concentration of 3.5 mg/ml is optimal for in vitro embryo attachment and trophoblastic cell outgrowth. Raising the concentration above 3.5 mg/ml does not improve embryo culture and can at certain concentrations be detrimental to embryo development.     

The modulus of elasticity of lobster aorta microfibrils

The presence of elastic fibres in the extracellular matrix (ECM) provides physiologically important elastic properties for many tissues. Until recently, microfibrils, one component of the ECM, were thought primarily to serve as a scaffolding on which elastin is deposited during development to form elaunin fibres [1]. The most prominent protein that forms mammalian microfibrils is fibrillin. It is known that mutations in the fibrillin gene cause a heterogenous connective tissue disease called marfan syndrome [2], so information on mechanical properties of microfibrils or their role in tissue function would be useful. Microfibrils are also found in the ECM of some invertebrate tissues, and there is growing evidence that the protein forming the structure is homologous to mammalian fibrillin [3, 4]. It has been shown that the microfibril-based arterial wall of the lobster has viscoelastic properties [5], and we have now utilized this primitive artery to measure the modulus of elasticity of microfibrils. It is similar to that of the rubber-like protein elastin.     

Extracellular microfibrils in development and disease

Fibrillins are the structural components of extracellular microfibrils that impart physical properties to tissues, alone or together with elastin as elastic fibers. Genetic studies in mice have revealed that fibrillin-rich microfibrils are also involved in regulating developmental programs and homeostatic processes through the modulation of TGF-β/BMP signaling events. A new paradigm has thus emerged whereby the spatiotemporal organization of microfibrils dictates both the cellular activities and physical properties of connective tissues. These observations have paved the way to novel therapeutic approaches aimed at counteracting the life-threatening complications in human conditions caused by dysfunctions of fibrillin-rich microfibrils. Received 2 April 2007; received after revision 23 May 2007; accepted 24 May 2007     

The use of 5-fluorocytosine and ketoconazole in the culture of the erythrocytic stages ofPlasmodium falciparum and some tumor cell lines

Summary In vitro culture systems are often contaminated by bacteria and fungi. It is therefore often necessary to supplement culture media with agents such as penicillin/streptomycin, gentamycin or amphotericin B. The latter cannot be used in the in vitro culture of erythrocytic stages ofP. falciparum, and thus anti-fungal agents have not been regularly used in this system. We describe the prophylactic use of 5-fluorocytosine (5-FC) and ketoconazole (KTZ) in tissue cultures at concentrations up to 300 and 10 g/ml respectively which have no effect on the growth ofP. falciparum (FCR-3 strain). A melanoma cell line (C32) and a line of uterine carcinoma (C41) were also unaffected by similar concentrations of 5-FC and KTZ. When dissolved in complete culture medium (RPMI 1640) with 10% human plasma, the minimum inhibitory concentration of 5-FC for a susceptible strain ofCandida remained below 2 g/ml. These experiments suggest that 5-FC (at 50 g/ml) alone or in combination with KTZ (at 1 g/ml) is a useful addition to the armamentarium of antimicrobials available to the tissue culture biologist for a variety of cell culture systems.     

Nutritional requirements of aeromonads and their multiplication in drinking water

Aeromonads can utilize a wide range of low molecular-weight compounds, including amino acids, carbohydrates and long-chain fatty acids at a concentration of a few micrograms per liter. Utilization of biopolymers such as gelatin, casein and amylose is slow at this concentration level. The concentration of substrates available for an A. hydrophila strain in drinking water was usually below 10 micrograms of C/l. The autochthonous bacteria utilized these substrates more rapidly than the aeromonads. The multiplication of aeromonads in drinking water during distribution is therefore explained by their growth on biomass components in the biofilm and in sediments in the pipes.     

加替沙星-壳聚糖纳米粒胶联羊膜药膜的制备及体外实验研究

目的制备一种新型生物多功能复合材料——加替沙星．壳聚糖纳米粒胶联羊膜药膜,并观察其体外缓释性能及抑菌作用。方法采用离子胶联法制备加替沙星－壳聚糖纳米粒并检测其表征,然后将栽药纳米粒加入到纤维蛋白胶中,混匀后再与羊膜胶联制成加替沙星－壳聚糖纳米粒胶联羊膜药膜,光镜及扫描电镜下观察其形态学特征,高效液相色谱法测定其中加替沙星的浓度,微量肉汤稀释法检测其体外抑菌活性。结果加替沙星纳米粒的粒径为291±33nm,载药量为10．46％,包封率为61．32％。载有纳米药物的纤维蛋白胶与羊膜基底面粘合紧密,呈圆形的加替沙星纳米粒附着在胶原纤维的网状结构上。在体外,加替沙星－壳聚糖纳米粒胶联羊膜药膜可持续释药达15天,对金黄色葡萄球菌和铜绿假单胞菌的最低抑菌浓度分别为0．05μg／ml和0．42μg／ml。结论加替沙星－壳聚糖纳米粒胶联羊膜药膜在体外具有良好的缓释性能及抑茵作用。     

Elastase, collagenase and the radial elastic properties of arteries

Studies were performed on 203 pairs of dog carotid arteries subjected to unidirectional radial compression. Treatment with 80 U/ml purified elastase for 90 min decreased radial stress, but treatment with 640 U/ml collagenase for 90 min did not. These data suggest that elastin, but not collagen, contributes to wall resistance to radial compression.     

Elastase,collagenase and the radial elastic properties of arteries

Summary Studies were performed on 203 pairs of dog carotid arteries subjected to unidirectional radial compression. Treatment with 80 U/ml purified elastase for 90 min decreased radial stress, but treatment with 640 U/ml collagenase for 90 min did not. These data suggest that elastin, but not collagen, contributes to wall resistance to radial compression.     

Effect of aloe lectin on deoxyribonucleic acid synthesis in baby hamster kidney cells

A homogeneous glycoprotein (mol. wt 40,000) containing 34% carbohydrate was isolated from Aloe arborescens var. natalensis. At a concentration of 5 micrograms/ml, this glycoprotein was shown to stimulate deoxyribonucleic acid (DNA) synthesis in baby hamster kidney (BHK) cells and to have the properties of a lectin which reacts with sheep blood cells. The chemical and physical properties of the glycoprotein (aloe lectin) are also discussed.     

Effect of actinomycin D on Trypanosoma cruzi

Viable metacyclic forms of T. cruzi, Y strain, treated with an adequate dose of actinomycin D (50 micrograms Act-D/ml/10(7) parasites/ml for 72 h at 28 degrees C) showed the following properties: 1) they lost their ability to replicate in culture medium, in blood and in tissues of normal mice and were no longer able to incorporate tritiated thymidine; 2) they could not penetrate into Vero cells and could not replicate inside normal macrophages; 3) they retained their immunogenicity and the ability to protect mice against a virulent infection; 4) they did not induce histological lesions as described in chronic experimental Chagas' disease.     

Mechanical and biochemical characterization of the contraction elicited by a calcium-independent myosin light chain kinase in chemically skinned smooth muscle

The contraction induced by a Ca2+-independent myosin light chain kinase (MLCK-) was characterized in terms of isometric force (Fo), immediate elastic recoil (SE), unloaded shortening velocity (Vus), shortening under a constant load and ATPase activity of chemically skinned smooth muscle preparations. These parameters were compared to those measured in a Ca2+ -induced contraction to assess the nature of cross bridge interaction in the MLCK-induced contraction. Fo developed in chicken gizzard fibers as well as SE were similar in contractions elicited by either agent. Vus in the contraction induced by MLCK-(0.36 mg/ml) was similar though averaged 39.3 +/- 8.9% less than Vus induced by Ca2+ (1.6 X 10(-6) M) in the control fibers. Addition of Ca2+ (1.6 X 10(-6) M) to a contraction induced by MLCK-resulted in small increases in both Fo and Vus. Shortening under a constant load was similar for both types of contractions. The contraction induced by MLCK-was accompanied by an increased rate of ATP hydrolysis. The MLCK-induced contraction is thus kinetically similar though not identical to a contraction induced by Ca2+. We conclude that with respect to actin-myosin interaction, MLCK-and Ca2+ -induced contractions are similar.     

The effect of actinomycin D on the production of viruses by the protoplasts of Chinese cabbage infected in vitro by the yellow mosaic virus of turnips]

Chinese Cabbage (Brassica sinensis L. var. Cantonner) protoplasts were infected by Turnip yellow mosaic virus (TYMV) and inoculated in the presence or absence of actinomycin D. Virus production was determined 40 hrs. after inoculation, the time required for the virus replication cycle to be terminated. While actinomycin D had no effect on TYMV production when present at a concentration of 1 microng/ml, a 50 to 80% inhibition of virus production was noticed at concentrations of the order of 5 to 10 microng/ml, and the inhibition reached 90% with 25 microng/ml.     

The effect ofβ-aminopropionitrile (BAPN) on bacteria

Zusammenfassung Alle untersuchten Bakterien waren gegenüber BAPN entweder ganz unempfindlich oder zeigten nur eine unspezifische Wachstumsverminderung. Diese Bakterien scheinen zum Studium lathyrogenischer Substanzen ungeeignet. Die Annahme, dass BAPN auf Mucopolysacchariden wirkt, konnte durch diese Untersuchung nicht gestützt werden.

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This work was supported by Research Grant A-4002 from the National Institute of Arthritis and Metabolic Diseases, U.S. Public Health Service. We are also grateful to Abbott Laboratories, North Chicago (Illinois) for generous gifts of mono--aminopropionitrile (BAPN) fumarate.