Does capsaicin have therapeutic benefits in human colon adenocarcinoma? Selection of the most reliable dose via AgNOR

Background/aim: To determine the effect of different doses of capsaicin on AgNOR protein synthesis in human colon adenocarcinoma derivate from colon cancer (Caco-2 cell). Materials and methods: In this experimental study, after the cultured of Caco-2 cell line, the cells are divided into 4 groups as control and different capsaicin exposed doses (25uµ, 50uµ, and 75uµ). Mean AgNOR number and total AgNOR area/nuclear area (TAA/NA) were calculated. Results: A significant differences were detected between control and capsaicin (50uµ) (P = 0.001), between control and capsaicin (75uµ) (P = 0.000), between capsaicin (25uµ) and capsaicin (50uµ) (P = 0.001) and between capsaicin (25uµ) and capsaicin (75uµ) (P = 0.000) for TAA/NA. Also, there were significant differences between control and capsaicin (50uµ) (P = 0.001), between control and capsaicin (75uµ) (P = 0.000), between capsaicin (25uµ) and capsaicin (50uµ) (P = 0.000) and between capsaicin (25uµ) and capsaicin (75uµ) (P = 0.000) for mean AgNOR number. Conclusion: A certain amount of capsaicin has a protective effect against colon adenocarcinoma and the dose concentrations are important for the most reliable treatment.

Effects of aluminum on nucleoli in root tip cells and selected physiological and biochemical characters in Allium cepa var. agrogarum L.

BACKGROUND: Increased Al concentration causes reduction of mitotic activity, induction of nucleolar alteration, increase of the production of ROS and alteration of several antioxidant enzyme activities in plant cells. Allium cepa is an excellent plant and a useful biomarker for environmental monitoring. Limited information is available about the effects of Al on nucleoli, antioxidant enzyme system, contents of MDA and soluble protein in A. cepa. Therefore, we carried out the investigation in order to better understand the effects of Al on the growth, nucleoli in root tip cells and selected physiological and biochemical characters. RESULTS: The results showed that the root growth exposed to 50 µM Al was inhibited significantly. 50 µM Al could induce some particles of argyrophilic proteins scattered in the nuclei and extruded from the nucleoli into the cytoplasm. The nucleolus did not disaggregate normally and still remained its characteristic structure during metaphase. Nucleolar reconstruction was inhibited. 50 µM Al induced high activities of SOD and POD in leaves and roots significantly (P < 0.05) when compared with control, whereas the level of CAT was low significantly (P < 0.05). At 50 µM Al the content of MDA in leaves was high significantly (P < 0.05) at 9(th) day and in roots increased (P < 0.05) with prolonging the treatment time during 6-12 days. The soluble protein content in leaves treated with 50 µM Al was high significantly (P < 0.05) at 6(th) day and increased with prolonging the treatment time. CONCLUSIONS: We suggest that variations in nucleoli and the alterations of antioxidant enzyme activities, MDA and soluble protein contents in Allium cepa can serve as useful biomarkers, which can provide valuable information for monitoring and forecasting effects of exposure to Al in real scenarios conditions. Among the antioxidant enzymes SOD and POD appear to play a key role in the antioxidant defense mechanism under Al toxicity condition. Data from MDA concentration show that Al indirectly produces superoxide radicals, resulting in increased lipid peroxidative products and oxidative stress.

[Effect of shenqi fuzheng injection on pre/post-operational change of argyrophilic-nucleolar organizer regions in peripheral T-lymphocyte in patients with gastric carcinoma].

OBJECTIVE: To investigate the pre-post operational change of argyrophilic-nucleolar organizer regions (Ag-NORs) in peripheral T-lymphocyte of patients with gastric carcinoma (GC), and to explore the effect of shenqi fuzheng injection(SFI) on it. METHODS: Eighty five patients were divided into two groups according to the operation performed was radical or non-radical, and the two groups were subdivided into two by additional intravenous dripping of SFI was given to them or not. The content of Ag-NORs in peripheral T-lymphocyte in all patients before and after operation as well as in 12 healthy subjects was determined. RESULTS: Content of Ag-NORs in GC patients was significantly lower than that in the healthy subject (P < 0.01), which significantly increased after patients underwent radical operation (P < 0. 01 or P < 0.05), especially in those treated with SFI (P < 0.01). While in patients underwent non-radical operation but not treated with SFI, it showed insignificant change after operation, however it did significantly increase in those treated with SFI (P < 0.05). CONCLUSION: The immune function of T lymphocyte was low in patients with gastric carcinoma, post-operational adjuvant treatment of SFI can significantly improve the cellular immunity of patients.

Staining human lymphocytes and onion root cell nuclei with madder root.

We performed staining experiments on cells using natural dyes and different mordants using techniques that are used for wool and silk dyeing. The natural dye sources were madder root, daisy, corn cockle and yellow weed. Ferrous sulfate, copper sulfate, potassium tartrate, urea, potassium aluminum sulfate and potassium dichromate were used as mordants. Distilled water, distilled water plus ethanol, heptane, and distilled water plus methanol were used as solvents. All dye-mordant-solvent combinations were studied at pH 2.4, 3.2 and 4.2. The generic staining procedure was to boil 5-10 onion roots or stimulated human lymphocyte (SHL) preparations in a dye bath on a hot plate. Cells were examined at every half hour. For multicolor staining, madder-dyed lymphocytes were decolorized, then stained with Giemsa. The AgNOR technique was performed following the decolorization of Giemsa stained lymphocytes. Good results were obtained for both onion root cells and lymphocytes that were boiled for 3 h in a dye bath that included 4 g madder root, 4 g ferrous sulfate as mordant in 50 ml of 1:1 (v/v) methanol:distilled water. The pH was adjusted to 4.2 with 6 ml acetic acid. We conclude that madder root has potential as an alternative dye for staining biological materials.

Modulation of carcinogenic response and antioxidant enzymes of rats administered with 1,2-dimethylhydrazine by Picroliv.

The effect of Picroliv treatment on the carcinogenic response and, hepatic and renal antioxidant enzymes of rats administered with 1,2-dimethylhydrazine hydrochloride (DMH) was studied in male Sprague-Dawley rats. DMH-induced hepatic carcinogenic response and necrosis were inhibited by oral administration of Picroliv (40 and 200 mg/kg). Liver gamma-glutamyl transpeptidase, which was elevated to 0.41 +/- 0.06 nmol/mg protein by DMH administration was found to be reduced to 0.22 +/- 0.04 and 0.18 +/- 0.03 nmol/mg protein by Picroliv treatment 40 and 200 mg/kg, respectively. Elevated number of Argyrophilic Nucleolar Organizer Region dots and clusters, an index of proliferation, of DMH treated rat liver was reduced by Picroliv treatment. DMH-induced depletion of hepatic and renal antioxidant enzymes such as catalase and superoxide dismutase levels were restored to normal by Picroliv treatment. Picroliv treatment reduced the DMH-induced elevation of lipidperoxidation in liver, kidney and serum. Elevated levels of serum total bilirubin by DMH administration was reduced by Picroliv treatment. Depleted renal glutathione S-transferase and hepatic glutathione levels after DMH administered rats were found to be significantly increased by Picroliv treatment. Histological analysis of the DMH administered rat liver showed hepatic cell necrosis, coalescent nodular areas and cystic hyperplasia of the bile ducts with inflammation. Picroliv treated liver resembled normal liver except the presence of a few degenerating cells. Renal anatomy was not altered by DMH administration.

Chemopreventive effects of scordinin on diethylnitrosamine and phenobarbital-induced hepatocarcinogenesis in male F344 rats.

Modifying effects of scordinin, a biological active component in garlic, on diethylnitrosamine (DEN)- and phenobarbital (PB)-induced hepatocarcinogenesis were examined in rats. Male F344 rats, 5 weeks old, were divided into 8 groups. After a week, groups 1 - 5 were given DEN (100 mg / kg body weight, i.p.) once a week for 3 weeks, whereas groups 6 - 8 received vehicle treatment. Group 2 was given 600 ppm scordinin-containing diet in the initiation phase. From 4 weeks after the start of experiment, groups 3 and 5 were fed scordinin, and groups 1 - 3 and 7 received drinking water containing 500 ppm PB. Group 6 was given scordinin diet alone throughout the experiment (24 weeks). The incidences of hepatocellular adenoma and carcinoma were significantly smaller in group 3 than those in group 1 (P < 0.005 and P < 0.05, respectively). The average numbers of liver neoplasms in groups 2 and 3 were significantly smaller than in group 1 (P < 0.01 and P < 0.0001, respectively). Glutathione S-transferase placental form-positive foci were also significantly decreased by scordinin treatment in the initiation or promotion phase. Scordinin significantly decreased the mean number of nucleolar organizer regions' protein (AgNORs) / nucleus in hepatocellular adenoma and carcinoma. AgNORs / nucleus in the non-lesional area was also significantly decreased by scordinin treatment during the promotion phase. These results suggest that scordinin is a promising chemopreventive agent for liver neoplasia.

The acetylation patterns of histones H3 and H4 along Vicia faba chromosomes are different.

The acetylation pattern of H3 was studied on field bean chromosomes by means of indirect immunofluorescence using polyclonal antibodies recognizing H3 isoforms acetylated at lysine positions 9/18, 14 and 23. H3 was found to be hypoacetylated at lysine residues 9/18 and 14 within the heterochromatic regions composed of tandem repetitive Fok-I elements. Hyperacetylation of these residues was observed at the nucleolar organizing region (NOR) and in heterochromatic regions composed of repeats other than Fok-I elements. In contrast, H4 was underacetylated (H4.Ac5, 8, 12) or uniformly acetylated (H4.Ac16) at all heterochromatic regions, and acetylated above the average at all four lysines only within the NOR. Acetylation of lysine-23 of H3 was uniform, except for the NOR that showed no fluorescence. Inhibition of deacetylase during and after replication of heterochromatin by trichostatin A had no influence on the acetylation status of H3 but mediated an increase in acetylation of lysines 5, 12 and 16 of H4 above the average in the field bean heterochromatin. Thus, the chromosomal acetylation patterns of H4 and H3 of this species revealed common and divergent features. Whereas the acetylation level of H4 correlates well with the potential transcriptional activity and inversely with the time of replication of defined chromatin domains of Vicia faba, this is not generally true for H3.

Histone H4 acetylation in plant heterochromatin is altered during the cell cycle.

Using polyclonal antibodies directed against acetylated isoforms of histone H4 (H4 acetylated at lysine positions 5, 8, 12, 16 and H4 tetraacetylated), indirect immunofluorescence revealed hyperacetylation for all H4 variants at the nucleolus organizer region (NOR) of metaphase chromosomes of the field bean Vicia faba. The transcriptionally inactive and late-replicating heterochromatin regions proved to be hypoacetylated at lysine positions 5, 8 and 12. The remaining chromatin showed average fluorescence. These patterns were altered when deacetylase was blocked by exposure of root tip meristems to trichostatin A for more than 2 h prior to fixation. Under these conditions, all lysine positions, except lysine 8, appeared to be hyperacetylated at the NOR and in addition at the prominent heterochromatin domains. This observation represents a hitherto unique switch of histone acetylation pattern during the cell cycle. This is apparently caused by deposition of acetylated H4. Ac5, 12 and 16 or by acetylation directly after replication, which later on becomes reduced (H4.Ac16) or even reversed (H4.Ac5 and 12) by deacetylase before cells enter mitosis.

[Effect of kidney tonifying herbs on morphological changes of adrenal cortex in androgen-sterilized rats].

Young rats of 9-day old with testosterone propionate subcutaneously injected were used as an androgen sterilized rats (ASR) model. On 80th day, herbal extract of tonifying the Kidney was fed for 14 days, all animals were sacrificed by cardiac perfusion method on 100 days of ages. Morphological studies with light and electron microscope, immunohistochemical studies with argyrophilic nucleolar organiser region (AgNOR) and proliferating cell nucleolar antigen (PCNA) were used for observations on the morphological change of adrenal cortex. In control group (treated with testosterone propionate only) the reticular zone of the adrenal cortex extented widely in which cellular fatty drops, AgNOR and PCNA increased apparently. In animal treated with herbs, the width of the reticular zone, the numbers of AgNOR and PCNA reduced significantly to normal levels (P < 0.01, P < 0.01, P < 0.01) and numbers of fatty drops also decreased. It suggests that during inducing ovulation in ASR tonifying Kidney herbs simultaneously regulated the adrenal function besides the chief function on pituitary-ovary regulation.

[The effects of electroacupuncture treatment on nucleolar organizer regions of adrenal cortex in ovariectomized rats].

The present paper reports the morphometric analysis of nucleolar organizer regions (NORs) of the adrenal cortex in ovariectomized rats following electroacupuncture (EA) using argyrephil (Ag-NOR) method for NORs. Animals were divided into four groups, the control group (CT group, n = 4), the EA group (n = 3), the ovariectomized group (OV group, n = 4) and the ovariectomized electroacupuncture group, (OV+EA group, n = 7). The number of AgNORs of 100 cells from zona fasciculata of the adrenal cortex in each case of all groups was counted at random and the mean +/- SE (number/cell) in each group was calculated as follows: OV+EA group 2.71 +/- 0.26, OV group 1.62 +/- 0.15, EA group 1.21 +/- 0.04 and CT group 1.48 +/- 0.03. The mean of AgNORs in OV+EA group differed highly significantly from the other three groups (P < 0.01) tested by ANOVA and LSD method, No significant distinction was found among the OV group, EA group and CT group. Gross specimen examination showed that adrenal glands in OV+EA were significantly heavier than those in the other three groups (P < 0.01). Vaginal smears showed that a response like that of estrogen-induced with exfoliative cells appeared in the OV+EA group rats following EA. EA had no influence on the change of exfoliative cells in EA group. The results suggest that EA may promote the synthesis and secretion of the adrenal steroid hormones, the androgen of which will then be transformed into estrogen in other tissues, thus compensating the deficiency of estrogen induced by ovariectomy.