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At least 1,603 records · Page 89Linked to original sources
PubMed2005

[Comparison of the immunomodulatory effects of spore polysaccharides and broken spore polysaccharides isolated from Ganoderma lucidum on murine splenic lymphocytes and peritoneal macrophages in vitro].

OBJECTIVE: To compare the immunomodulatory effects of spore polysaccharides (Gl-SP) and broken spore polysaccharides (Gl-BSP) isolated from Ganoderma lucidum(Leyss et Fr.) Karst. on murine splenic lymphocytes and peritoneal macrophages in vitro. METHODS: Mixed lymphocyte culture reaction (MLR), lymphocyte proliferation in the presence or absence of mitogen, and the cytotoxic activity of splenic natural killer (NK) cells were detected with MTT assay in vitro. The percentage of phagocytosis of neutral red (NR) by mouse peritoneal macrophages was detected by colorimetric assay. Splenic T-lymphocyte subpopulations were measured with flow cytometry(FCM). IL-2, IFN-gamma and TNF-alpha in the culture supernatants were detected by ELISA and biological assay. Nitric oxide (NO) production was examined by Griess reaction. RESULTS: At the concentration range of 0.2-12.8 mg/L, Gl-SP and Gl-BSP were shown to increase lymphocyte proliferation in the presence or absence of mitogen, enhance NK cytotoxic activity, augment the production of TNF-alpha and NO in Gl-SP- or Gl-BSP-activated macrophages, as well the percentage of phagocytosis of NR by macrophages in vitro. Both Gl-SP and Gl-BSP could promote MLR, however, at the dose of 12.8 mg/L, Gl-BSP showed higher activity than Gl-SP in the proliferation of lymphocytes. These two kinds of polysaccharide could significantly increase the secretion of IL-2 and IFN-gamma in doublejway MLR at the concentrations of 0.2-12.8 mg/L, but Gl-BSP had stronger effects than Gl-SP at the same concentrations. Both Gl-SP and Gl-BSP could increase the ratio of T-lymphocyte subpopulations in double-way MLR. At the concentrations of 0.2-12.8 mg/L or 3.2-12.8 mg/L, Gl-BSP demonstrated more significant activity in increasing the percentage of the CD4(+) or CD8(+) subset than Gl-SP. At the concentrations of 0.2-0.8 mg/L, the ratio of the CD4(+) and CD8(+) subset in the Gl-BSP treated group was higher than that of the Gl-SP treated group. CONCLUSION: Gl-SP and Gl-BSP have similar immunomodulatory effects in vitro, as though the immunomodulatory effects of Gl-BSP are stronger than that of Gl-SP.

Adjuvants, Immunologic↗
PubMed2005

[Cytotoxicity of chemicals used in household products: 1997- 2004].

The cytotoxicities of chemicals used in household products were evaluated using a neutral red (NR) uptake assay. The chemicals tested during 1997-2004 were rubber additives (accelerators, antioxidants and retarders), solvents, plasticizers and biocides, such as antimicrobials, fungicides, preservatives used in paints, paper, wood and plastic products. The cytotoxicity potential of each chemical was classified by determining the concentrations inducing 50% reduction of NR uptake into Chinese hamster fibroblast V79 cells compared to control (IC50). In vivo eye irritancy of each chemical was estimated by the IC50 value. Most biocides tested showed strong cytotoxicity and had a high probability of inducing strong eye irritation.

Animals↗
PubMed1975

Photodynamic inactivation of herpesvirus hominins by methylene blue (38524).

Methylene blue, in a concentraion of 10-5M was virostatic in the presence of light but not in the dark for 31 of 37 strains of fresh isolates of herpesvirus hominis. Resistance to the dye developed during treatment. This photodynamic pattern was almost identical to that of neutral red which produced an identical effect. This was not ture for proflavine which was active in the dark in many instances. Cross insensitivity between proflavine and methylene blue was not observed.

Coloring Agents↗
PubMed2006

[Effects of traditional tibetan medicine, Fructus Lonicerae microphyllae on phagecytosis and cytokines production of murine macrophages].

OBJECTIVE: To explore the effects of traditional Tibetan medicine, Fructus Lonicerae microphyllae (FLM) on phagecytosis and cytokines production of murine macrophages. METHOD: The phagecytosis of murine macrophages was analyzed by neutral red phagecytosis assay. The activities of IL-1 and TNF-alpha were measured by biological methods. The mRNA of TNF-alpha and INF-gamma expressed by macrophages was detected by RT-PCR. RESULT: The phagecytosis of murine macrophages was significantly enhanced by FLM at a concentration from 1 microg x mL(-1) to 100 microg x mL(-1) and the secretions of IL-1, and TNF-alpha from macrophages were markedly induced by FLM. Meanwhile, FLM also increased the expression of TNF-alpha mRNA and INF-gamma mRHA from macrophages in vitro. CONCLUSION: FLM could promote phagecytosis and cytokines production of murine macrophages.

Animals↗
PubMed2006

[Effect of gingerol on endotoxemia mouse model induced by heatstroke].

OBJECTIVE: To observe the effect of Gingerol on endotoxemia mouse induced by heatstroke. METHODS: Forty mice were randomly divided into five groups, the endotoxemia model group (A), the normal temperature group (B), the Gingerol treated group (C), the solvent control group (D), and the saline control group (E), 8 mice in each group. Group B to E was administered with saline, Gingerol, solvent and saline respectively. Mice in group B were placed at room temperature 25 +/- 0.5 degrees C , relative humidity 43 +/- 5 % for 2 hrs, while mice in the other groups were exposed under 35 +/- 0.5 degrees C and relative humidity 65 +/- 5 % for 2 hrs in an artificial hot-climate mimic cabin to establish heatstroke endotoxemia model. The energy metabolic level of celiomacrophage was detected with MTT; the phagocytic ability was examined with neutral red chromometry; the hepatocyte ultrastructure was observed with transmission electron microscopy, as well as the activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in plasma was tested. RESULTS: As compared with Group A, D and E, in Group C, energy metabolic levels of macrophage, phagocytic ability, and activity of SOD were significantly higher (P < 0.01), and the level of MDA was significantly lower respectively (P < 0.01), with the levels of SOD and MDA approaching to those in Group B (P >0.05). The pathologic changes of hepatocyte ultrastructure in group C were less than those in the other three endotoxemia groups. CONCLUSION: Gingerol could raise the energy metabolic level of celio-macrophage to enhance its phagocytic ability, increase the activity of SOD and reduce the production of MDA in mouse with heatstroke endotoxemia, so as to alleviate the liver damage.

Animals↗
PubMedDate not supplied

Pharmacological and toxicological evaluation of a new series of thymidylate synthase inhibitors as anticancer agents.

Thymidylate synthase (TS) is responsible for catalysing the de novo biosynthesis of doexythymidine monophosphate and is a target for many anticancer drugs. A series of thymidylate synthase inhibitors (TSIs), synthesised in our laboratory, were submitted to primary anticancer screening by the National Cancer Institute (NCI). Four compounds, 3,3-bis(4-methoxyphenyl)-1H, 3H-naphtho[1,8-cd]pyran-1-one (MR7), 6-chloro-3,3-bis(4-hydroxyphenyl)-1H,3H-naphtho[1,8-cd]pyran-1-one (MR21), 3,3-bis(3-fluoro-4-hydroxyphenyl)-1H,3H-naphtho[1,8-cd]pyran-1-one (MR35) and 6-bromo-3,3-bis(3-chloro-4-hydroxyphenyl)-1H,3H-naphtho[1,8-cd]pyran-1-one (MR36), passed the criteria and were automatically scheduled for evaluation against the full panel of 60 human tumour cell lines. In this study, the antiproliferative activity of the substances against SK-MEL-2 cells (from metastatic tissue) and SK-MEL-28 cells (from primary malignant melanoma cells) was investigated. Neutral Red uptake and the MTT test were performed to confirm the results of the NCI, and [3H]-thymidine incorporation was performed as a test of the proliferation rate. Our results indicated that compounds MR21 and MR36 were the most active agents and the [3H]-thymidine test was the best in predicting toxicity against melanoma cells.

Antineoplastic Agents↗
PubMed2006

[Effects of TCE and PCE on cultured human keratinocyte lipid peroxidation and protective effect of vitamin E on it].

OBJECTIVE: To explore the different concentrations of trichloroethylene (TCE) or perchloroethylene (PCE) induced cultured normal human epidermal keratinocyte (KC) lipid peroxidation and protective effect of Vitamin E on it. METHODS: KC derived from 3 or more donors were pooled together and cultured with K-SFM. Neutral Red Uptake Assay was used to determine the IC50 of TCE or PCE, and then different concentrations of TCE or PCE were administered for culturing KC; 0.5 mmol/L TCE or 0.2 mmol/L PCE and different concentrations of Vitamin E were used to determine the protective effect of Vitamin E. After 4 hours' culture, kits were used to determine cellular MDA, SOD and ROS level. RESULTS: Treatment of KC with different concentrations of TCE or PCE showed significant dose-related variations in lipid peroxidation, with the higher concentration, higher level of MDA, ROS and lower activity of SOD displayed in this study. Vitamin E 10 - 200 mmol/L dose-dependently attenuated MDA and ROS level, and increased SOD activities. CONCLUSION: TCE or PCE can induce the lipid peroxidation in cultured KC and Vitamin E protects it from TCE- or PCE-induced peroxidation.

Antioxidants↗
PubMed1991

[Comparative analysis of the dimensions of vestibulo-spinal neurons in the guinea pig].

The comparative analysis of dimensions of horseradish peroxidase labelled vestibulo-spinal neurons and neutral red stained neurons of undifferentiated areas of the vestibular nuclei have been performed in guinea pig. The undifferentiated areas of the vestibular nuclei consisted of small, medium, large and giant cells. A decrease in the relative numbers of small- and medium-diameter neurons and an increase in the relative numbers of large and giant neurons have been observed along the caudo-rostral axis of the vestibular complex. The vestibulo-spinal populations included neurons of medium, large and giant dimensions. The numerical superiority of large and giant neurons have been found. The even distribution of the vestibulo-spinal neurons of different diameters have been revealed along the longitudinal axis of the brainstem.

Animals↗
PubMed1976

Recurrent conjugal neuralgia caused by Herpesvirus hominis type 2.

Leg neuralgia is usually caused by nerve root irritation, and is seldom considered of viral origin. Two married couples had recurrent leg pain in a dermatome distribution. In each episode, after one to three days of neuralgia, herpetic vesicles appear on or near the genitalia. Herpesvirus hominis (HVH) was isolated from vesicles or cervix in three patients and was serologically identified as herpesvirus honinis type 2. Prolonged follow-up has not shown progression of the syndrome or any permanent neurologic damage. Treatment with topical neutral red and photoinactivation has not only reduced local lesion healing time and frequency of attacks, but it has also given prompt relief of neuralgia.

Adult↗
PubMed1991

Areca-nut toxicity in cultured human buccal epithelial cells.

In cultured human buccal epithelial cells, at doses of 3-540 micrograms/ml, areca-nut extract significantly decreased viability, as determined by colony-forming efficiency, clonal growth rate, ability to take up neutral red and ability to exclude trypan blue, and also caused significant formation of DNA single-strand breaks and DNA protein cross-links. Comparisons of the areca nut-related compounds, 3-(N-nitrosomethylamino)propion-aldehyde (NMPA), 3-(N-nitrosomethylamino)propionitrile (NMPN), N-nitrosoguvacoline, N-nitrosoguvacine, arecoline, arecaidine, guvacoline and guvacine, in terms of the above endpoints, indicate that NMPA is ten times more cytopathic to buccal cells than the other agents on a molar basis. Because metabolism of NMPA can potentially yield several reactive breakdown products, including aldehydes, this study indicates that both the parent compound and its metabolites may contribute to the observed pathobiological effects. Taken together, the observed pathobiological effects of areca-nut extract and certain related compounds in cultured human buccal epithelial cells indicate that these agents may contribute to the oral carcinogenicity associated with chewing betel quid.

Areca↗
PubMed1976

[Stick-plaque test--an economic method of quantitative determination of viruses].

An economic method for quantitative assay of viruses is presented. In this "canule stick-plaque test" (German abbreviation SPT) samples of viruses, geometrically diluted and taken up by a canule, are inoculated by sticking into monolayer cell cultures overlayed with agar medium. A plaquelike CPE detectable by neutral red staining develops in the area of the inoculation. The frequency of this CPE formation depends on the concentration of viruses in the inoculated dilution. This dose-response allows calculation of the ID50. In this way it is possible to carry out titration involving 6 dilutions and 10 inoculations per dilution using 3 common Petri dishes (6 cm in diameter), only. The sensitivity, accuracy, and reproductibility of this method are described and discussed.

Cells, Cultured↗
PubMed1976

[Stich-plaque test--an economic method for quantitative determination of viruses].

An economic method for quantitative assay of viruses is presented. In this "canule stick-plaque test" (German abbreviation SPT) samples of viruses, geometrically diluted and taken up by a canule, are inoculated by sticking into monolayer cell cultures overlayed with agar medium. A plaquelike CPE detectable by neutral red staining develops in the area of the inoculation. The frequency of this CPE formation depends on the concentration of viruses in the inoculated dilution. This dose-response allows calculation of the ID 50. In this way it is possible to carry out titration involving 6 dilutions and 10 inoculations per dilution using 3 common Petri dishes (6 cm in diameter), only. The sensitivity , accuracy, and reproductibility of this method are described and discussed.

Animals↗
PubMed1991

Toxic effects of interleukin-2-activated lymphocytes on vascular endothelial cells.

IL-2-activated lymphocytes (LAK cells) show increased adherence to, and killing of, human vascular endothelial cells compared to resting lymphocytes. In the present work, we have found that supernatants from LAK cell cultures also are toxic to human umbilical vein endothelial cells (HUVEC) when tested for 48 h in a neutral red uptake assay. Recombinant TNF-alpha and IFN-gamma at high concentrations are also toxic under the same test conditions, and TNF-alpha was directly detected in LAK cell supernatants. An inconsistent inhibition of toxicity was found with anti-TNF-alpha whereas anti IFN-gamma antibodies had a partial inhibitory effect. The susceptibility of HUVEC to cellular killing by LAK cells could be up- and down-regulated with insulin-like growth factor I and IFN-gamma, respectively. It is concluded that damage to vascular endothelium during high dose IL-2 treatments may be partially related to an excessive production of lymphokines such as IFN-gamma and TNF-alpha. IFN-gamma may, in addition, be protective for HUVEC during cellular interactions with LAK cells.

Culture Media↗
PubMed1991

[Intravital determination of pH gradient between the cytoplasm and lysosomes in human fibroblasts in the normal state and in hereditary storage diseases].

Presented are the results of measurements of pH in cytoplasm and lysosomes of skin fibroblasts of healthy donors and patients with lysosomal storage diseases, mannosidosis, Fabry, Krabbe disease. The pH value was estimated in the stationary phase of growth using neutral red (lysosomes) and fluorescein diacetate (cytoplasm). It was shown that the cytoplasmic pH value in pathological cells didn't virtually differ from the control values. The intralysosomal pH value in fibroblasts of patients with mannosidosis and Fabry disease was essentially increased, which correlated with the size increase of these organelles upon the accumulation of unsplit compounds. This led to the decrease in pH gradient between the cytoplasm and lysosomes in the pathological cells, an increase in intralysosomal pH along with hereditary deficiency of enzymes could bring about the retardation of catabolic processes in lysosomes.

Cells, Cultured↗
PubMed1991

Modification of leech behavior patterns by reserpine-induced amine depletion.

A single injection of 100 micrograms reserpine into the crop of the medicinal leech, Hirudo medicinalis, reduced CNS serotonin and dopamine levels to less than 1% of control values within 3 d. High-pressure liquid chromotography- (HPLC) determined CNS serotonin and dopamine levels remained maximally depressed for approximately 1 month following reserpine injection. Subsequently, amine levels recovered slowly, but remained depressed 6 months after reserpine injection. Following reserpine treatment, glyoxylic acid-induced fluorescence or neutral red staining closely mirrored the HPLC-determined time course of amine depletion and recovery. Acute exposure of isolated ganglia to 10 microM reserpine for periods up to 6 hr produced a 20-30% reduction of serotonin and dopamine content. The threshold concentration of reserpine necessary to produce amine depletion was approximately 1 microM. We found that reserpine treatment eliminated biting behavior within 4 d following injection. Biting behavior remained depressed below control levels for approximately 4 months, but returned to control values while CNS serotonin and dopamine levels remained significantly depressed at this time. Unexpectedly, reserpine treatment increased rather than reduced the duration of stimulus-evoked swimming activity. This behavioral change was evident within 3 d and persisted for approximately 3.5 months. To rapidly restore amine levels in reserpine-treated animals, we bathed intact leeches in pond water containing serotonin, dopamine, or octopamine. We found that biting behavior was restored following reserpine treatment by bathing intact leeches in pond water containing serotonin or dopamine, but not octopamine. Also contrary to expectations, the increase in swim duration was not reversed by bath exposure to serotonin, dopamine, octopamine, or histamine. However, all swimming activity in reserpine-treated leeches was eliminated by the amine antagonist cyproheptadine. We propose that the presence of low levels of amines is critical for the expression of both biting and swimming activity in leeches. However, the minimal levels of amines necessary for the expression of these behaviors are lower for swimming than for biting.

Animals↗
PubMed1978

Cultural, morphological, cell membrane, enzymatic, and neoplastic properties of cell lines derived from a Hodgkin's disease lymph node.

A neoplastic cell line (designated HuT11) has been established in continuous culture from an involved lymph node of a patient with Stage IIA Hodgkin's disease of the mixed cellularity type. The HuT11 line has been morphologically heterogeneous, consisting of mononucleate lymphoid-like cells, polygonal epithelioid cells, and mono-, bi-, and multinucleate giant cells. Four clones initiated from isolated binucleate giant cells of the HuT11 line also have been successfully established as continuous cell lines. The cloned lines have been morphologically distinct and more homogeneous, although typical giant cells have consistently appeared throughout the long-term culture of each. The HuT11 lines have grown as monolayers in McCoy's Medium 5A supplemented with 10% fetal calf serum, with generation times of 12 to 14 hr and high saturation densities. Cytogenetic studies showed that early and later passages of HuT11 cells were aneuploid, and all cell lines were successfully heterotransplanted in the hamster cheek pouch. Repeated indirect immunofluorescence examinations have shown each cell line to be negative for Epstein-Barr virus nuclear antigen. Indirect immunofluorescence tests in which monospecific immunoglobulins were used revealed positive membrane reactions for the gamma (heavy)-chain and kappa (light)-chain of human immunoglobulin G in approximately 20% of viable cells in each line; however, direct immunofluorescence with anti-human immunoglobulin G F(ab')2 reagent failed to confirm these reactions. Rosette tests for B- and T-lymphocyte and macrophage membrane receptors yielded negative results. All cell lines were strongly phagocytic for latex particles and neutral red dye. Cytochemical stains of the monolayers revealed abundant esterase, fluoride-resistant nonspecific esterase, acid phosphatase, and leucine aminopeptidase activities, while lysozyme assays were negative. Although some properties of the HuT11 lines have suggested a macrophage derivation, an undifferentiated lymphoid cell origin of the Hodgkin's neoplastic cell remains a possibility.

Aneuploidy↗
PubMed1990

In vitro toxicity of lomefloxacin in rabbit corneal epithelial cell cultures.

Lomefloxacin (NY-198) is a chemotherapic agent from the new 4-quinolone group, acting on DNA gyrase system. Lomefloxacin (LFLX), as some other new 4-quinolone compounds are antimicrobials of potential use for ophthalmic application. A first approach on the oculotoxicity of LFLX is the main purpose of this study. Four concentrations of LFLX (10, 30, 100 and 300 micrograms/ml) were tested on the first subculture of pigmented rabbit corneal epithelial cells. Cell number, protein contents, neutral red stain and wound healing were evaluated. The results showed that the lower concentrations (10 & 30 micrograms/ml) had no effect while the highest one had a remarkable cytotoxic effect. Pharmacokinetic data show that peak values achieved in the cornea and other ocular structures are lower than 40 micrograms/g (0.3% topical application) and, on another hand, MIC values range from less than or equal to 0.05 to 16 micrograms/ml. So, these "critical" concentrations have no cytotoxic effects according to our results. Then, it is concluded that Lomefloxacin could be a useful drug for topical ophthalmic development.

4-Quinolones↗
PubMed1990

[A pH increase in human fibroblast lysosomes during accumulation of non-hydrolysable compounds].

The changes in intralysosomal pH were measured in the stationary phase of normal human embryonic fibroblast growth under sucrose loading over a period of 6 to 120 hours and in cells with a typical lysosomal storage pathology, Fabry's disease, using a vital indicator dye, neutral red. It was shown that long-term hypertrophy of the lysosomal compartment during intracellular accumulation of non-hydrolysable compounds is concomitant with a pH increase, on the average, by 0.4 units. The highest values of pH (7.0-7.2) were seen in large-sized heterogeneic lysosomes of pathological cells. It is suggested that an increase in intralysosomal pH during accumulation of non-hydrolysable compounds leads to deterioration of conditions that are favourable for the acidic hydrolase function.

Cells, Cultured↗