Leukocyte Nucleolus and Anisakis pegreffii-When Falling Apart Means Falling in Place.

The view of the nucleolus as a mere ribosomal factory has been recently expanded, highlighting its essential role in immune and stress-related signalling and orchestrating. It has been shown that the nucleolus structure, formed around nucleolus organiser regions (NORs) and attributed Cajal bodies, is prone to disassembly and reassembly correlated to various physiological and pathological stimuli. To evaluate the effect of parasite stimulus on the structure of the leukocyte nucleolus, we exposed rat peripheral blood mononuclear cells (PBMC) to the crude extract of the nematode A. pegreffii (Anisakidae), and compared the observed changes to the effect of control (RPMI-1640 media), immunosuppressive (MPA) and immunostimulant treatment (bacterial lipopolysaccharide (LPS) and viral analogue polyinosinic:polycytidylic acid (poly I:C)) by confocal microscopy. Poly I:C triggered the most accentuated changes such as nucleolar fragmentation and structural unravelling, LPS induced nucleolus thickening reminiscent of cell activation, while MPA induced disassembly of dense fibrillar and granular components. A. pegreffii crude extract triggered nucleolar segregation, expectedly more enhanced in treatment with a higher dose. This is the first evidence that leukocyte nucleoli already undergo structural changes 12 h post-parasitic stimuli, although these are likely to subside after successful cell activation.

Quantitative analysis of AgNOR counts and pap stain in exfoliative cytology specimens of oral mucosa in bidi smokers and nonsmokers.

Background: Bidi smoking is a serious health hazard which is common throughout South Asia and parts of the Middle East. It has been strongly implicated to various benign and malignant lesions of oral cavity and oropharynx. These tobacco-filled leaves deliver more than three times the amount of nicotine, carbon monoxide, and tar as cigarettes which exert injurious effects on cells reflected in terms of accelerated proliferative activity in normal oral mucosal cells. Aim: This study aimed to compare the exfoliated cells from the oral mucosa of bidi smokers and nonsmokers, with emphasis on proliferative activity. Materials and Methods: Exfoliative smears were obtained from the oral mucosa of forty participants (twenty nonsmokers and twenty smokers) with age group ranging from 30-80 years, in and around Barwala (Haryana). The cytologic smears were evaluated using Papanicolaou (PAP) stain and AgNOR in order to evaluate the presence of cytological alterations suggestive of inflammation, dysplasia, keratinization, and proliferative activity of epithelial cells. Only PAP Class I and Class II smears were observed. Results: Comparison of the mean number of AgNORs showed a significant difference between nonsmokers and smokers. Inflammatory alterations were found in 70% of smokers and 55% of nonsmokers. A significant difference in proliferative activity was observed between smokers and nonsmokers classified as PAP Class II. Conclusion: A significant difference of AgNORs/nucleus was observed between bidi smokers and nonsmokers.

RésuméContexte: Bidi fumeurs est un grave danger pour la santé qui est commune dans toute l'Asie du Sud et certaines parties du Moyen-Orient. Il a été fortement impliqué dans diverses lésions bénignes et malignes de la cavité buccale et l'oropharynx. Ces feuilles de tabac offrent plus de trois fois la quantité de nicotine, monoxyde de carbone et de goudron que les cigarettes qui exercent des effets préjudiciables sur les cellules reflétés sous la forme d' une accélération de l'activité proliférative des cellules de la muqueuse buccale normale. Objectif: Cette étude visait à comparer les cellules exfoliées de la muqueuse orale des bidis fumeurs et non fumeurs, avec l'accent sur l'activité proliférative. Matériels et méthodes: frottis Exfoliative ont été obtenus à partir de la muqueuse orale de 40 participants (20 non-fumeurs et fumeurs) avec 20 Groupe d'âge allant de 30-80 ans, dans et autour de Barwala (Haryana). Le frottis cytologique ont été évalués à l'aide de la coloration de Papanicolaou (PAP) et d'AgNOR afin d'évaluer la présence d' altérations cytologiques évocateurs d'infl ammation, dysplasie, la kératinisation, et l'activité proliférative des cellules épithéliales. PAP uniquement les catégories I et II de Papanicolaou n'a été observé. Résultats: comparaison du nombre moyen d'AgNORs ont montré une différence entre les non-fumeurs et les fumeurs. Des modifications ont été trouvés infl ammatory dans 70% des fumeurs et 55% des non-fumeurs. Une différence dans l'activité proliférative a été observée entre les fumeurs et les non-fumeurs PAP, le niveau d'emploi de la classe II. Conclusion: une différence de AgNORs/noyau a été observée entre fumeurs et non-fumeurs bidi. Mots-clés: AgNOR, frottis exfoliative, les fumeurs.

Decreased argyrophilic nucleolar organiser region (AgNOR) expression in Purkinje cells: first signal of neuronal damage in sudden fetal and infant death.

AIMS: The nucleolus is an important cellular component involved in the biogenesis of the ribosome. This study was performed in order to validate the introduction of the argyrophilic nucleolar organiser region (AgNOR) stain technique, specific for the nucleoli detection, in neuropathological studies on sudden fetal and infant death. METHODS: In a wide set of fetuses and infants, aged from 27 gestational weeks to eight postnatal months and dead from both known and unknown causes, an in-depth neuropathological study usually applied at the Lino Rossi Research Center of the Milan University was implemented by the AgNOR method. RESULTS: Peculiar abnormalities of the nucleoli, as partial or total disruption above all in Purkinje cells (PCs), were exclusively found in victims of sudden fetal and infant death, and not in controls. The observed nucleolar alterations were frequently related to nicotine absorption in pregnancy. CONCLUSIONS: We conclude that these findings represent early hallmarks of PC degeneration, contributing to the pathophysiology of sudden perinatal death.

Are there any effects of chronic carbon monoxide exposure on argyrophilic nucleolar-organizing region-associated protein synthesis in rat myocardium?

The aims of the study are to detect whether there are any possible effects of chronic carbon monoxide (CO) exposure on the argyrophilic nucleolar-organizing region (AgNOR)-associated protein synthesis and evaluate any possible relationship between the amount of AgNOR protein and the level of myocardial injury also and between AgNOR and histopathological evaluation methods. Adult male albino Wistar rats (n = 18) were randomly divided into three groups (groups A, B, and C). Group A served as control, while groups B and C were rats exposed to CO gas chronically (1000 and 3000 ppm CO concentration with a flow rate of 4 L/min for 30 min/day for 7 days, respectively). Total AgNOR area/nuclear area (TAA/NA) and the mean AgNOR numbers for each myocyte nucleus were determined. There were significant differences among all groups for TAA/NA ratio. These differences were not significant for mean AgNOR numbers. According to the histopathological evaluation scores, there were significant differences between the groups. The differences were significant among the groups for loss of sarcomere pattern. A strong positive correlation between histopathological injury scores and TAA/NA ratio was found (Rsq = 0.48; p = 0.002), however, the correlation was not significant for mean AgNOR numbers (Rsq = 0.08; p = 0.25). In conclusion, TAA/NA ratio can be used as an indicator for obtaining information about the level of myocardial damage instead of histopathological evaluation scores.

Effect of Hydroxyapatite Nanoparticles on the Growth Potential of Hepatoma Cells in Nude Mice.

AIM: To evaluate the activity of hydroxyapatite (HAP) nanoparticles against the proliferation of hepatoma cells. METHODS: HAP nanoparticles were prepared by homogeneous precipitation. The size distribution and morphology of these nanoparticles were determined by laser particle analysis and transmission electron microscopy, respectively. Xenograft tumor models of human hepatoma cells (Bel-7402) implanted in nude mice under the right scruff skin were established and divided into two groups: treatment and control. Once the xenograft tumor grew to a diameter of 0.8 cm, 0.2 ml HAP nanoparticle suspension was injected into the tumor every day for 2 weeks. The long and short diameters of the tumors were measured before and after HAP injection, and the inhibition rate of tumor growth was calculated. Paraffin tissue sections were prepared from xenograft tumors treated as above for 2 weeks, histologically stained for DNA and agyrophilic nucleolar organizer region (AgNORs), and immuno-histologically stained for proliferating cell nuclear antigens (PCNAs). The stained sections were examined by microscopy. Images of these sections were recorded and analyzed by image analysis system and relevant software for DNA content, AgNOR intensity, and PCNA expression in the nucleus, nucleoli, and hepatoma cells, respectively. RESULTS: The HAP nanoparticles were uniformly distributed, with a size of 44.6 nm to 86.8 nm. Upon the local injection of the tumor with the HAP nanoparticles, the average volumes of the tumors were significantly reduced compared with those of the control group, which had a tumor inhibition rate of 51.32%. The DNA content, AgNOR intensity, and PCNA expression in the hepatoma cells were all significantly reduced (P < 0.01) compared with those in the control group. CONCLUSION: HAP nanoparticles inhibit the proliferation of hepatoma cells in vivo.

Ellagic Acid-Changed Epigenome of Ribosomal Genes and Condensed RPA194-Positive Regions of Nucleoli in Tumour Cells.

We studied the effect of ellagic acid (EA) on the morphology of nucleoli and on the pattern of major proteins of the nucleolus. After EA treatment of HeLa cells, we observed condensation of nucleoli as documented by the pattern of argyrophilic nucleolar organizer regions (AgNORs). EA also induced condensation of RPA194-positive nucleolar regions, but no morphological changes were observed in nucleolar compartments positive for UBF1/2 proteins or fibrillarin. Studied morphological changes induced by EA were compared with the morphology of control, non-treated cells and with pronounced condensation of all nucleolar domains caused by actinomycin D (ACT-D) treatment. Similarly as ACT-D, but in a lesser extent, EA induced an increased number of 53BP1-positive DNA lesions. However, the main marker of DNA lesions, γH2AX, was not accumulated in body-like nuclear structures. An increased level of γH2AX was found by immunofluorescence and Western blots only after EA treatment. Intriguingly, the levels of fibrillarin, UBF1/2 and γH2AX were increased at the promoters of ribosomal genes, while 53BP1 and CARM1 levels were decreased by EA treatment at these genomic regions. In the entire genome, EA reduced H3R17 dimethylation. Taken together, ellagic acid is capable of significantly changing the nucleolar morphology and protein levels inside the nucleolus.

Curcumin-mediated decrease in the expression of nucleolar organizer regions in cervical cancer (HeLa) cells.

Curcumin, the major yellow-orange pigment of turmeric derived from the rhizome of Curcuma longa, is a highly pleiotropic molecule with the potential to modulate inflammation, oxidative stress, cell survival, cell secretion, homeostasis and proliferation. Curcumin, at relatively high concentrations, was repeatedly reported to be a potent inducer of apoptosis in cancer cells and thus considered a promising anticancer agent. In the present paper, the effects of low concentrations of curcumin on human cervical cancer (HeLa) cells were studied. We found curcumin-mediated decrease in the cell number and viability, and increase in apoptotic events and superoxide level. In contrast to previously shown curcumin cytotoxicity toward different cervical cancer lines, we observed toxic effects when even as low as 1 µM concentration of curcumin was used. Curcumin was not genotoxic to HeLa cells. Because argyrophilic nucleolar protein (AgNOR protein) expression is elevated in malignant cells compared to normal cells reflecting the rapidity of cancer cell proliferation, we evaluated curcumin-associated changes in size (area) and number of silver deposits. We showed curcumin-induced decrease in AgNOR protein pools, which may be mediated by global DNA hypermethylation observed after low concentration curcumin treatment. In summary, we have shown for the first time that curcumin at low micromolar range may be effective against HeLa cells, which may have implications for curcumin-based treatment of cervical cancer in humans.

[Effect of peptide bioregulator and cobalt ions on the activity of NORs and associations of acrocentric chromosomes in lymphocytes of patients with hypertrophic cardiomyopathy and their relatives].

The influence of peptide bioregulator - Livagen (Lys-Glu-Asp-Ala) separately and combined with cobalt ions, on the activity of nucleolar organizer regions (NORs) and frequency of associations of acrocentric chromosomes in lymphocytes from patients with hypertrophic cardiomyopathy (HCM) and their relatives has been studied. It is shown that combined action of Livagen and cobalt ions increases the frequency of large-sized scoring 2 NORs in both, patients and their relatives. Significant was also the influence of the studied compounds on associative activity of acrocsentric chromosomes that was expressed in sharp increase of this indicator in both studied groups. In this case more effective was the action of Livagen and cobalt ions. As activity of NOR, also the frequency of associations of acrocentric chromosomes is dependent of quality of acrocentric chromosome stalk condensation, we conclude, that by influence of Livagen and cobalt ions on the lymphocytes of HCM patients and their relatives, occurs decondensation of heterochromatinized chromatin. This may be release condition during condensation of inactivated genes in the studied groups of individuals. Our data are important because it provides new information about protective effect of Livagen and Livagen+Cobalt ions on the lymphocytes of HCM patients and their relatives and may lead to the development of a therapeutic treatment.

Identification of novel markers that demarcate the nucleolus during severe stress and chemotherapeutic treatment.

The nucleolus, the ribosomal factory of the cell, has emerged as a key player that regulates many aspects of cell biology. Several thousand proteins associate at least transiently with nucleoli, thereby generating a highly dynamic compartment with a protein profile which is sensitive to changes in cell physiology and pharmacological agents. Powerful tools that reliably demarcate the nucleoli are a prerequisite to measure their composition and activities. Previously, we developed quantitative methods to measure fluorescently labeled molecules in nucleoli. While these tools identify nucleoli under control and mild stress conditions, the accurate detection of nucleolar boundaries under harsh experimental conditions is complicated by the lack of appropriate markers for the nucleolar compartment. Using fluorescence microscopy we have now identified new marker proteins to detect nucleoli upon (a) severe stress and (b) drug treatments that trigger a pronounced reorganization of nucleoli. Our results demonstrate that nucleolin is an ideal marker to delimit nucleoli when cells are exposed to heat or oxidative stress. Furthermore, we show for the first time that cellular apoptosis susceptibility protein (CAS) and human antigen R protein (HuR) are excluded from nucleoli and can be employed to delimit these compartments under severe conditions that redistribute major nucleolar proteins. As proof-of-principle, we used these markers to demarcate nucleoli in cells treated with pharmacological compounds that disrupt the nucleolar organization. Furthermore, to gain new insights into the biology of the nucleolus, we applied our protocols and quantified stress- and drug-induced changes in nucleolar organization and function. Finally, we show that CAS, HuR and nucleolin not only identify nucleoli in optical sections, but are also suitable to demarcate the nucleolar border following 3D reconstruction. Taken together, our studies present novel marker proteins that delimit nucleoli with high confidence under a variety of experimental settings.

[Inhibitory effects of doxycycline on argyrophilic nucleolar organizing regions and a-smooth muscle actin expression in proliferative bovine corneal myofibroblasts in vitro].

OBJECTIVE: To investigate the effect of doxycycline on the nucleolar organizing regions and a-smooth muscle actin expression in bovine corneal myofibroblasts in vitro and assess its contribution to ocular surface repair mechanisms. METHODS: Cell culture and identification: bovine corneal fibroblasts were cultured after the stroma was incubated in 1.0 and 2.0 g/L type I collagenase in two stages.Isolated cells were plated at mantaryay culture flask in 10% of BSA RPMI-1640. Vimentin and alpha-smooth muscle actin (α-SMA) organization were evaluated by immunocytochemistry. The cells staining positive for Vimentin and α-SMA indicated the presence of corneal myofibroblasts. Bovine corneal myofibroblasts were treated with different concentrations of doxycycline (10, 20, 40, 60, 80 mg/L) , a bland control group and the dexamethasone group (120 mg/L) were set up, each group had 30 cases. The argyrophilic nucleolar organizing regions (AgNOR) staining and the immunohistochemistry for α-SMA were performed when the cells were treated for 24 hours and 48 hours. The AgNOR count (Ag-c), AgNOR area (Ag-a) and the expression of α-SMA in the bovine corneal myofibroblasts among each experiment group and control group were compared using one-way ANOVA, further pairwise comparisons using Independent-Samples t test. RESULTS: Cell culture techniques were successfully used to establish a method for the isolation and culture of bovine corneal myofibroblasts. Microscopic examination and immunohistochemical staining confirmed that the cells cultured were bovine corneal myofibroblasts. The Ag-c and Ag-a of bovine corneal myofibroblasts progressively decreased as the concentrations of doxycycline was increase. 24 h:bland control group Ag-c was 6.40 ± 0.6, 60 mg/L doxycycline group Ag-c was 2.23 ± 0.43;bland control group Ag-a was (34.80 ± 2.36) µm(2), 60 mg/L doxycycline hormone group Ag-a was (19.91 ± 2.15) µm(2). 48 h: bland control group Ag-c was 7.27 ± 0.6,60 mg/L doxycycline hormone group Ag-c was 2.80 ± 0.76, bland control group Ag-a was (36.27 ± 1.99) µm(2), 60 mg/L doxycycline group Ag-a was (13.75 ± 2.09) µm(2). The differences were statistically significant: in the same time intervention (FAg-c 24 h = 252.55, FAg-a 24 h = 202.16, P < 0.05, FAg-c 48 h = 169.38, FAg-a 48 h = 853.23, P < 0.05), in the same concentrations intervention (tAg-c = 6.98, tAg-a = 11.62, P < 0.05). And 60 mg/L of doxycycline had an obviously inhibitory action as 120 mg/L dexamethasone in the same treated hours (dexamethasone group Ag-a 24 h = 30.56 ± 3.66, dexamethasone group Ag-a 48 h = 28.35 ± 1.23 ),the differences were not statistically significant (tAg-a 24h = 1.182, P = 0.242,tAg-a 48 h = 0.21, P = 0.832). As the concentrations investigated, doxycycline can inhibit the expression of α-SMA in the bovine corneal myofibroblasts (189.90 ± 7.48, 140.20 ± 7.79, 113.20 ± 8.98, 98.00 ± 3.50, 85.50 ± 4.99), the difference was statistically significant (F = 761.79, P = 0.00). While dexamethasone had no significant role in the expression of α-SMA (bland control group was 225.10 ± 6.74, the dexamethasone group was 228.50 ± 7.12), and the statistically difference was not obvious (t = 1.096, P = 0.287). CONCLUSIONS: As the concentrations of doxycycline was increased from 10 mg/L to 80 mg/L, the AgNOR count and AgNOR area of bovine corneal myofibroblasts can be significantly reduced in vitro. Compared with dexamethasone, doxycycline significantly suppressed the expression of α-SMA in bovine corneal myofibroblasts in a dose-dependent positive trend.

Exfoliative cytological assessment of apparently normal buccal mucosa among quid chewers using argyrophilic nucleolar organizer region counts and Papanicolaou staining.

OBJECTIVE: Quid chewing is associated with an increased risk of oral cancer. This study aims to analyze argyrophilic nucleolar organizer region (AgNOR) counts along with Papanicolaou (PAP) staining in exfoliative smears of quid chewers and non-chewers to correlate quid chewing habits with possible early cytological changes in apparently normal buccal mucosa. STUDY DESIGN: Exfoliative smears were obtained from normal buccal mucosa of 30 male quid chewers and non-chewers. The smears were stained using the AgNOR silver staining technique to evaluate the proliferative activity and PAP for cytological appearance. RESULTS: Statistically higher AgNOR counts were observed in chewers as compared to non-chewers. The difference in the mean percentage of nuclei having ≥5 AgNORs in both groups was statistically significant (p = 0.001). In chewers, PAP showed 77% with class I and the remaining 23% were class II, while the non-chewers showed only class I cytology. AgNOR counts between chewers and non-chewers having class I cytological appearance demonstrated a greater mean AgNOR count in chewers (p = 0.0001). CONCLUSION: Quid chewing seems to have a definite role in promoting proliferative activity of apparently normal buccal mucosal cells. Exfoliative cytological assessment of a combination of AgNOR counts and PAP has the potential for prediction of early quid-associated cellular changes before the appearance of clinical premalignant and malignant lesions.

Antiproliferative efficacy of hesperetin (citrus flavanoid) in 1,2-dimethylhydrazine-induced colon cancer.

Cancer is the second leading cause of death worldwide and is increasing at an alarming rate. The present study was to evaluate the antiproliferative effects of hesperetin, a flavonoid commonly found in many herbal medicines and foods, on aberrant crypt foci (ACF), argyrophylic nucleolar organizer regions (AgNORs) and proliferating cell nuclear antigen (PCNA) in 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in rats. Rats were given subcutaneous injections of DMH (20 mg/kg body weight) weekly for 15 weeks to induce carcinogenesis, and hesperetin was administered orally at the dose of 20 mg/kg body weight. DMH exposure alone produced a high incidence of ACF and showed positive staining for PCNA and AgNORs in colonic tissues. Supplementation with hesperetin lowered the PCNA labeling index and suppressed the formation of ACF in the rats with colon cancer. These results clearly reveal that dietary hesperetin possesses antiproliferative ability against chemically induced colon tumourigenesis.

[Deheterochromatinization of the chromatin in old age induced by oligopeptide bioregulator (Lys-Glu-Asp-Pro)].

In this work is presented the data on the variability of the functional characteristics of the chromosomes in the cells exposed by oligopeptide bioregulator - Prostamax from old individuals (75-86 years). Evaluated: the frequency of sister chromatid exchanges (SCE); Ag-positive NORs (in associations and nonassociations), as well as the variability of the structural C-pericentromeric heterochromatin. Prostamax changed the chromosomal parameters: 1) increased the frequency of SCE to 12,0±0,28 exchange in per cell (in intact cells - 5,9±0,2); 2) increased the frequency of Ag-positive NORs to 2.5 per cell (in intact cells - 0.95) 3) reduced in the frequency of large segments of the options from the pericentromeric heterochromatin for the 1st and 9th chromosomes. Comparison of the results indicates the ability of Prostamax to decondensation, deheterchromatinization the chromatin during aging, and thus release by heterochromatinization repressed genes. On the other hand, the data obtained in this work suggest that the basis for the protective action of Prostamax its modifying effect on chromatin.

The nucleolus directly regulates p53 export and degradation.

The correlation between stress-induced nucleolar disruption and abrogation of p53 degradation is evident after a wide variety of cellular stresses. This link may be caused by steps in p53 regulation occurring in nucleoli, as suggested by some biochemical evidence. Alternatively, nucleolar disruption also causes redistribution of nucleolar proteins, potentially altering their interactions with p53 and/or MDM2. This raises the fundamental question of whether the nucleolus controls p53 directly, i.e., as a site where p53 regulatory processes occur, or indirectly, i.e., by determining the cellular localization of p53/MDM2-interacting factors. In this work, transport experiments based on heterokaryons, photobleaching, and micronucleation demonstrate that p53 regulatory events are directly regulated by nucleoli and are dependent on intact nucleolar structure and function. Subcellular fractionation and nucleolar isolation revealed a distribution of ubiquitylated p53 that supports these findings. In addition, our results indicate that p53 is exported by two pathways: one stress sensitive and one stress insensitive, the latter being regulated by activities present in the nucleolus.

Evaluation of hyaluronan gel (Gengigel(®) ) as a topical applicant in the treatment of gingivitis.

AIM: To clinically and histopathologically evaluate the anti-inflammatory effect of 0.2% hyaluronan gel alone and with mechanical therapy on gingivitis. The argyrophilic nucleolar organizer region staining technique was attempted to routinely determine its diagnostic and prognostic dependability for periodontal lesions. METHODS: In each of the 28 gingivitis patients, the four quadrants were subjected to different treatments: scaling, scaling + topical hyaluronan gel, only topical hyaluronan gel, and topical + intrasulcular hyaluronan gel. Clinical parameters were recorded at baseline, and on days 7, 14, and 21. Biopsies were taken from each quadrant, inflammatory infiltrates were graded, and the argyrophilic nucleolar organizer region count was measured before and after treatment. RESULTS: A significant reduction was seen in clinical parameters, inflammatory infiltrates, and the argyrophilic nucleolar organizer region count within the groups. The effect of topical + intrasulcular gel was equivalent to scaling (P > 0.05). Topical + intrasulcular hyaluronan gel application demonstrated a better reduction than topical hyaluronan gel alone. CONCLUSIONS: Hyaluronan gel is an effective topical agent for treating gingivitis, along with scaling and intrasulcular application. The argyrophilic nucleolar organizer region count can be used as a histopathological indicator in cases of non-responsive gingivitis to assess the severity of gingival inflammation.

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 finasteride on serum prostate-specific antigen (PSA) and on prostate of hamster Mesocricetus auratus (hMa).

PURPOSE: Evaluate the effects of finasteride on the serum PSA and on the prostate of hamster-Mesocricetus auratus(hMa). METHODS: Twenty hMa male adults were split in groups control and experimental (n=10). Animals of the experimental group received 7.14ng/mL of finasteride, subcutaneously (SC) on the back three times per week, during 90 days. The finasteride dose was equivalent to 5.0mg administered to a 70kg man. At the end of the experiment the mean age for the animals in the control group was 15.2 + or - 1.13 months and for the experimental group was 17.7 + or - 0.67 months. There was a statistically significant difference between mean ages of both groups (t value=5.98; p=0.001). The animals of the control group weighted 129.0 + or - 18.8g and the experimental group weighted 145.0 + or - 15.5g, t=1.88 e p=0.0514. The serum PSA was assessed through ELISA method. Prostates of those animals were collected and processed to histology and morphometry: the diameter of the acinous glands and the acinous epithelium, apoptosis, AgNORs and cellularity were assessed in both groups. RESULTS: Serum PSA decreased in the experimental group, 0.003ng/mL versus 0.763ng/mL, H= 7.982 e p= 0.0047. Decrease in the acinous area occurred in animals that received finasteride, 238.000 + or - 24.600 microm(2) versus 398.600 + or - 55.320 microm(2); t= 2.653; p= 0.0122. A remarkable decrease in the area of the acinous epithelium occurred in the animals that received finasteride, 111.900 + or -12.820 microm(2) versus 160.400 + or - 18.430 microm(2) t= 2.162; p= 0.0361. AgNORs were less expressed in finasteride treated animals, 2.846 + or - 0.877 versus 3.68 + or - 1.07 argyrophilic clusters for microm(2), p= < 0.0001. Apoptosis was more intense in the experimental group, 53.62 + or - 1.389 than in controls, 14.76 + or - 2.137, p= 0.0408. However, there was no statistical difference in the cellularity between both groups, 74.75 + or - 5.5 cells, in controls versus 65.07 + or - 13.24, in treated animals, p=0.5105. CONCLUSIONS: Use of finasteride decreased serum PSA and several histological parameters of the Hamster's prostate, such as lumen area, acinous and epithelium area, and AgNORs expression. Finasteride increased apoptosis in the prostate acinous cells although no significant difference in the cellularity could be found between the two groups of animals under study.

Effect of finasteride on serum prostate-specific antigen (PSA) and on prostate of hamster Mesocricetus auratus (hMa) / Efeito da finasterida no antígeno prostático específico (PSA) sérico e na próstata do hamster Mesocricetus auratus (hMa)

PURPOSE: Evaluate the effects of finasteride on the serum PSA and on the prostate of hamster-Mesocricetus auratus(hMa). METHODS: Twenty hMa male adults were split in groups control and experimental (n=10). Animals of the experimental group received 7.14ng/mL of finasteride, subcutaneously (SC) on the back three times per week, during 90 days. The finasteride dose was equivalent to 5.0mg administered to a 70kg man. At the end of the experiment the mean age for the animals in the control group was 15.2±1.13months and for the experimental group was 17.7±0.67 months. There was a statistically significant difference between mean ages of both groups (t value=5.98; p=0.001). The animals of the control group weighted 129.0±18.8g and the experimental group weighted 145.0±15.5g, t=1.88 e p=0.0514. The serum PSA was assessed through ELISA method. Prostates of those animals were collected and processed to histology and morphometry: the diameter of the acinous glands and the acinous epithelium, apoptosis, AgNORs and cellularity were assessed in both groups. RESULTS: Serum PSA decreased in the experimental group, 0.003ng/mL versus 0.763ng/mL, H= 7.982 e p= 0.0047. Decrease in the acinous area occurred in animals that received finasteride, 238.000±24.600 μm² versus 398.600±55.320 μm²; t= 2.653; p= 0.0122. A remarkable decrease in the area of the acinous epithelium occurred in the animals that received finasteride, 111.900±12.820 μm² versus 160.400±18.430 μm² t= 2.162; p= 0.0361. AgNORs were less expressed in finasteride treated animals, 2.846±0.877 versus 3.68 ±1.07 argyrophilic clusters for μm², p= < 0.0001. Apoptosis was more intense in the experimental group, 53.62±1.389 than in controls, 14.76 ± 2.137, p= 0.0408. However, there was no statistical difference in the cellularity between both groups, 74.75±5.5 cells, in controls versus 65.07±13.24, in treated animals, p=0.5105. CONCLUSIONS: Use of finasteride decreased serum ...

OBJETIVO: Avaliar o efeito da finasterida no PSA sérico e na próstata do hamster-Mesocricetus auratus (hMa). MÉTODOS: 20 hMa adultos machos foram divididos em grupos de 10 animais. No experimento foram administrados 7,14 ng/mL de finasterida, subcutâneo (SC), no dorso, três vezes por semana, por 90 dias, dose equivalente a 5,0 mg usada em homem de 70Kg. Ao final da pesquisa, grupo experimento apresentou idade média de 17,7 ± 0,67 meses. O grupo controle apresentou idade média de 15,2 ± 1,13 meses. O valor de t na comparação das médias das idades entre os dois grupos foi de 5,98 e p=0.0001. Os animais-controle pesaram em média 129,0 ± 18,8g e o experimento 145,0 ± 15,5g; t=1,88 e p=0,0514. Na microscopia óptica de luz e estudo morfométrico: avaliaram-se o diâmetro dos ácinos e epitélio acinar prostáticos, a apoptose, a expressão AgNORs e a celularidade. RESULTADOS: O grupo-experimento apresentou média de PSA de 0,003 ng/mL e o grupo-controle de 0,763 ng/mL, H=7,982 e p=0,0047. A área dos ácinos do grupo-experimento foi de 238,000±24,600 μm² versus 398,600±55,320 μm²; t= 2,653; p= 0,0122. A área do epitélio acinar no grupo-experimento foi de 111,900±12,820 μm² versus 160,400±18,430 μm² t= 2,162; p= 0,0361. A expressão de AgNORs foi menor no grupo-experimento 2,846±0,877 versus 3,68 ±1,07 grumos argilófilos por μm², p= < 0,0001. A apoptose foi mais freqüente no grupo-experimento, 53,62±1,389 versus controle, 14,76 ± 2,137, p= 0,0408. Não houve diferença na celularidade entre os grupos de animais, 74,75±5,5 células no grupo-controle versus 65,07±13,24, no grupo-experimento, p= 0,5105. CONCLUSÕES: A finasterida diminuiu o PSA sérico, a área do lúmen, o epitélio acinar, a expressão de AgNORs e promoveu a apoptose nos ácinos da próstata dos hamsteres experimento e não houve diferença na celularidade acinar entre os animais estudados.

[Cytogenetic effects of cholinotropic preparations mixture on Chironomus plumosus (Diptera) larvae in vivo].

Morphometric analysis of changes in nucleolar organizer (NO), Balbiani rings (BR)--BR(B), BR(1G), BR(2G) and chromosome I arm B puff activities, and in chromosome compactness of Chironomus plumosus (Diptera) polytene chromosomes was carried out in acute period under separate and combined influence of atropine and pilocarpine. Supression effect of cholinotropic preparations mixture was revealed. Suppression of NO activity with atropine concentration increase in the mixture served as criterion of toxicity.

Effect of indoor air pollution from biomass fuel use on argyrophilic nuclear organizer regions in buccal epithelial cells.

This study investigated the effect of indoor air pollution from biomass-fuel use on the expression of argyrophilic nucleolar organizer regions (AgNORs), an indicator of ribosome biosynthesis, in epithelial cells of oral mucosa. AgNORs were evaluated using cytochemical staining in 62 nonsmoking indian women (median age, 34 years), who cooked exclusively with biomass, and 55 age-matched women, who were from a similar neighborhood and cooked with relatively clean liquefied petroleum gas (LPG). Concentrations of particulate pollutants in indoor air were measured using a real-time aerosol monitor. Compared to the LPG-using controls, biomass-fuel users showed a remarkably increased number of AgNOR dots per nucleus (6.08 +/-2.26 vs 3.16 +/-0.86, p < 0.001), AgNOR size (0.85 +/-0.19 vs 0.53 +/-0.15 mum2, p < 0.001), and percentage of AgNOR-occupied nuclear area (4.88 +/-1.49 vs 1.75 +/-0.13%, p < 0.001). Biomass-using households had 2 to 4 times more particulate pollutants than that of LPG-using households. The changes in AgNOR expression were positively associated with PM10 and PM2.5 levels in indoor air after controlling for potential confounders such as age, kitchen location, and family income. Thus, biomass smoke appears to be a risk factor for abnormal cell growth via upregulation of ribosome biogenesis.