Argyrophilic Nucleolar Organizer Region Staining Research Articles

Evaluation of the protective role of resveratrol against sepsis caused by LPS via TLR4/NF-κB/TNF-α signaling pathways: Experimental study.

The development and progression of sepsis are multifactorial and influencethe immunological, endocrine, and cardiovascular systems of the body. Our knowledge of the key mechanisms involved in the pathogenesis of sepsis has expanded exponentially, yet this still needs to be translated into effective targeted therapeutic regimes. In the present study, we aimed to determine whether resveratrol has positive effects in the experimental sepsis rat model. Twenty-eightmale Spraque-Dawley rats were randomly divided into four groups (n = 7) as follows:control, lipopolysaccharide (LPS) (30 mg/kg dose), resveratrol, and LPS and resveratrol. After the experiment, liver and kidney tissues were collected for histopathological evaluation, blood serums were collected to measure malondialdehyde levels with enyzme-linked immunosorbent assay, and Toll-like receptor-4 (TLR4), tumor necrosis factor-α (TNF-α), nuclear factor-κB (NF-κB) immunoreactivity density was evaluated immunohistochemically. In addition, messenger RNA expression levels for TLR4, TNF-α, NF-κB, interleukin-1β, and interleukin 6 were measured. In addition, the damage observed in liver and kidney tissue was determined by AgNOR (argyrophilic nucleolar organizer regions) staining. LPS application caused severe tissue damage, oxidative stress, and increased the expressions of proinflammatory proteins and genes we evaluated, while resveratrol application eliminated these negativities. Resveratrol has been proven to suppress the TLR4/NF-κB/TNF-α pathway, a possible therapeutic signaling pathway that is important in initiating the inflammatory response in an animal model of sepsis.

Adapted technique for demonstrating argyrophilic nucleolar organizer regions in the cytologic samples of canine mast cell tumors.

Argyrophilic nucleolar organizer regions (AgNORs) are nonhistone argyrophilic nucleolar proteins associated with ribosomal genes found in the nucleolar organizer region that reflect cell proliferation and have an affinity for silver. AgNOR staining may be useful to evaluate prognosis in several neoplasms because higher AgNOR counts are related to higher grade tumors, metastases, and shorter survival times. We aimed to report on a quick and practical technique to identify AgNORs adapted for use in routine cytology. The cytopathologic diagnosis of mast cell tumor (MCT) in samples collected by fine-needle aspiration (FNA) was determined. Next, slides were impregnated with a solution containing silver nitrate; the main modification of our technique included incubation of these slides at a controlled temperature of 25 °C. Some slides were previously stained with Diff-Quik and others were only fixed with methanol. The slides were analyzed under a microscope, and the number of blackened intranuclear points (AgNORs) was counted. Slides prestained with Diff-Quik were easily counted compared with slides only fixed in methanol. Technical issues encountered with the methanol-fixed slides included insufficient cellularity, background precipitation, and an absence of silver impregnation. The technique reported in this study showed satisfactory results for AgNOR counting in cytologic smears from MCT, such as good impregnation and the elimination of background interferents. Further evaluation of this method comparing AgNOR counts with histologic examinations, tumor grades, other prognostic markers, and survival times are needed to fully evaluate the benefit of this technique.

Evaluation of antitumoral effect of mistletoe fruit extract on Ehrlich ascites tumor cells with muse cell analyzer and argyrophilic nucleolar organizer region staining method

Abstract Introduction Mistletoe has been used alone or as a complementary therapy in the treatment of different diseases for years. In this study, the antitumoral effect of mistletoe fruit extract on Ehrlich ascites tumor (EAT) cells was evaluated. Materials and Methods EAT cells from preformed stock mice were transferred to culture dishes containing 5-fluorouracil (5-FU) and mistletoe extracts at different doses (100, 200, 400, and 800 μg/ml). These cells were incubated at 37 °C in an environment with 95% humidity and 5% CO2. At the end of the incubations, the apoptosis status of the cells, cell cycle, mitochondrial membrane potential, and proliferation status with the argyrophilic (Ag) nucleolar organizer region staining (NORs) method were evaluated. Results As a result, it was observed that the mistletoe fruit extract and 5-FU induce apoptosis of EAT cells. It was concluded that the 5-FU substance arrests the cell cycle at the G0/G1 stage, while the mistletoe arrests the cell cycle at the S and G2/M stages. The depolarization rate of the mistletoe treated cells was higher. As a result of the evaluation made with the AgNORs method, it was seen that mistletoe and 5-FU could be effective in reducing the proliferation of EAT cells. Conclusions It was seen that mistletoe fruit extract could be effective in stimulating the apoptosis and depolarization of cancer cells. The results of other studies in the literature and our study support each other. It was concluded that the mistletoe plant may be useful in cancer treatment.

Avaliação quantitativa da expressão da proteína PCNA e das AgNORs em ameloblastomas

In order to assess the proliferative behaviour of the ameloblastoma epithelium nine cases of this neoplasm were retrieved from the files of the Oral Pathology Service laboratory of Rio Grande do Sul Federal University School of Dentistry. Two proliferation marker techniques were employed: the AgNor - argyrophilic nucleolar organizer regions staining and the proliferation cell nuclear antigen (PCNA) immunohistochemical assessment technique. Each case was submitted to a quantitative and comparative statistical analysis of the results obtained for the two techniques. The conclusion arrived at shows ameloblastoma presenting benign behavior, an average AgNOR index of 1.612, and a PCNA proteindependent proliferative activity with average immunoreactivity of 51.61%. The histological patterns analyses of the neoplastic epithelial component demonstrates that cells resembling pre-ameloblasts have obtained figures statistically higher to those obtained for cells resembling the stellate reticulum of the enamel organ, in both techniques employed. Therefore, it might be inferred that cells resembling pre-ameloblasts hold the proliferative potential of ameloblastomas and are probably responsible for the growth characteristics and infiltrative ability of this tumour.

The Correlate between Argyrophilic Nucleolar Organizer Region Stain and Ki-67 Immnochistochemistry in Diagnostic Breast Cancer

Introduction: This experimental study gauged the value of argyrophilic nucleolar organizer region (AgNOR) staining as a possible technique for the estimation of cell kinetics in conventional histology sections, in benign and malignant breast lesions. Methods: With a silver staining technique and immunohistochemistry, we associated the numbers of AgNORs and Ki67 scores in 30 breast carcinomas and 10 benign breast lesions. Results: The mean values of Ag NORs silver stain dots count for normal, benign, grade II and III were 1.28±0.17, 2.83±0.68, 5.23±0.87 and 7.32±0.92, respectively. There was a statistically significant difference (p≤0.001) was noticed between the all individual groups, among the normal and breast lesion as well as among the GII and GIII. Immunohistochemical Results of Ki-67 protein exhibited homogenous golden-brown color in control case and a positive brown granules or diffuse dark brown color in the nuclei of both benign and malignant cases under the 400X magnify examined under the light microscope. Discussion: AgNOR counts performed on routine formalin-fixed paraffin sections could provide substantial kinetic evidence. Additionally, the difference in AgNOR counts between benign and malignant tumors is such that they may be of diagnostic worth.

Argyrophilic nucleolar organizer regions in oral mucosa cells of crack and cocaine users: a cross-sectional study.

The objective of this study was to analyze and compare the presence of argyrophilic nucleolar organizer regions (AgNORs) in exfoliated cells obtained from the buccal mucosa among crack and cocaine users and non-users by the AgNOR staining technique. Seventy-three males ≥ 18years were categorized into two groups: 38 crack and/or cocaine users and 35 non-user volunteers. They were interviewed and responded a questionnaire regarding general health and drug addiction. Exfoliative cytology specimens were collected from the clinically intact buccal mucosa, and cytological preparations were fixed and stained by AgNOR technique. The mean number of AgNORs (p = 0.02) and the percentage of epithelial cells with more than 3 (p = 0.01) and 4 (p = 0.04) AgNORs/nucleus were significantly higher in the non-user group. In conclusion, the frequency and diversity of substances present in the drugs-cocaine, crack, and alcohol-consumed by the volunteers of this study may have influenced the number of AgNORs and the response to damage and consequent effect on protein synthesis.

Argyrophilic nucleolar organizer regions staining for cytology smears in dogs and cats.

The argyrophilic nucleolar organizer regions (AgNORs) are cellular proliferation markers, crucial for predicting the clinical course and aggressiveness of tumors. The purpose of this study was to establish an easy and practical AgNOR staining method in the cytology of dogs and cats. Air-dried cytological slides were prepared from dogs (n=14) and cats (n=12). Acetone, formalin, ethanol and methanol were tested as fixatives for AgNOR staining. Subsequently, various methods of Romanowsky-based counterstains were tested before and after AgNOR staining. Clear and strong AgNOR spots were observed with all fixatives, and post-May–Grünwald staining was the best counterstaining method. The established method showed clear AgNOR spots even in the long-term storage samples and Romanowsky-stained ones.

Periodontal implications of argyrophilic nucleolar organizer region: A mini reviewPeriodontal implications of argyrophilic nucleolar organizer region: A mini review

The periodontal lesions with cellular proliferation can be assessed by various methods. One of the methods to determine the proliferative activity is silver-staining argyrophilic nucleolar organizer region (AgNOR) staining. AgNOR can be used as diagnostic and prognostic tool for many of periodontal lesions which are proliferative in nature and may be quantitative marker of incipient cellular alterations before the histologic hallmarks appear. Hence, this paper reviewed about the periodontal applications of AgNOR staining.

Role of Argyrophilic Nucleolar Organiser Region Staining in Effusions for Detection of Malignancy - Diagnostic Test Evaluation

Role of Argyrophilic Nucleolar Organiser Region Staining in Effusions for Detection of Malignancy - Diagnostic Test Evaluation

In vitro growth and development of isolated secondary follicles from vitrified caprine ovarian cortex.

The aim of this study was to evaluate the viability, antrum formation and in vitro development of isolated secondary follicles from vitrified caprine ovarian cortex in a medium previously established for fresh isolated secondary follicles, in the absence (α-minimum essential medium (α-MEM+) alone) or presence of FSH and vascular endothelial growth factor (VEGF; α-MEM++FSH+VEGF). Ovarian fragments were distributed among five treatments (T1 to T5): fresh follicles were fixed immediately (T1), follicles from fresh tissue were cultured in vitro in α-MEM+ (T2) or α-MEM++FSH+VEGF (T3) and follicles from vitrified tissue were cultured in vitro in α-MEM+ (T4) or α-MEM++FSH+VEGF (T5). After 6 days of culture, treated follicles (T2, T3, T4 and T5) were evaluated for morphology, viability and follicular development (growth, antrum formation and proliferation of granulosa cells by Ki67 and argyrophilic nucleolar organiser region (AgNOR) staining). The levels of reactive oxygen species (ROS) in the culture media were also assessed. Overall, morphology of vitrified follicles was altered (P<0.05) compared with the fresh follicles. Follicular viability, antrum formation and ROS were similar between treatments (P>0.05). The average overall and daily follicular growth was highest (P<0.05) in T3. Granulosa cells in all treatments (T1, T2, T3, T4 and T5) stained positive for Ki67. However, fresh follicles from T3 had significantly higher AgNOR staining (P<0.05) compared with follicles of T1, T2, T4 and T5. In conclusion, secondary follicles can be isolated from vitrified and warmed ovarian cortex and survive and form an antrum when growing in an in vitro culture for 6 days.

Quantitative analysis of Argyrophilic Nucleolar organizer regions in odontogenic cysts and tumor – A comparative study

BackgroundThe nucleolar organizer region (NOR) is by definition part of a chromosome, and nucleolus is a structure containing this chromosomal part and in addition the material which accumulate around the NOR, mostly rRNAs and their precursors as well as specific ribosomal proteins. Argyrophilic Nucleolar organizing region (AgNOR) are silver binding NORs often used to study cell proliferation in various types of tumors. AimQuantitative assessment of Argyrophilic Nucleolar organizing region count and its comparison among dentigerous cyst, keratocystic odontogenic tumor and ameloblastoma. Material and methodForty-five histologically confirmed cases, 15 cases each of keratocystic odontogenic tumor, dentigerous cysts and ameloblastomas were examined for Argyrophilic Nucleolar organizing region. The sections were obtained and Argyrophilic Nucleolar organizer regions staining was done for comparing the proliferative capacity among these lesions. ResultPost hoc analysis for inter-group comparison and one way ANOVA were done in all three groups in this study. P<0.001 was considered significant. The results of AgNOR counts were higher in KCOTs as compared to ameloblastoma and least in dentigerous cysts. The mean AgNOR counts between the study groups were compared using one way ANOVA test and the differences were found to be significant (P<0.001). ConclusionAgNOR counts were significantly higher in KCOT and ameloblastoma as compared to dentigerous cyst suggesting that these lesions have a higher proliferative capacity than dentigerous cyst. The finding of a significantly higher AgNOR counts in KCOT as compared to ameloblastoma represent a difference in proliferative activity and greater growth potential between these two lesions.

Prognostic potential of AgNORs in oral submucous fibrosis

Aim and Objective:The role of prognosis cannot be stressed enough, especially when it comes to potentially malignant lesions. The argyrophilic nucleolar organizer regions (AgNORs), which is simple and cost-effective has been used in diagnostic and prognostic pathologies. This study seeks to identify the nucleolar organizer regions (NORs) in oral submucous fibrosis (OSMF), to correlate the AgNOR count with the histologic grade of OSMF, and to evaluate the prognostic potential of AgNOR.Materials and Methods:The sample size consisted of archival paraffin blocks of 35 cases of varying grades of OSMF and 10 cases of squamous cell carcinoma. Normal mucosa samples served as controls for the study. AgNOR staining in accordance with the method of Smith and Crocker was performed and Student's t-test was used for statistical analysis.Results:The results showed an increase in AgNOR counts with corresponding grades of OSMF, the count being least in normal mucosa and also an increase in AgNOR count with corresponding decrease in differentiation of oral squamous cell carcinoma.Conclusion:AgNOR staining is a rapid and inexpensive procedure representing cellular proliferation that can be used to assess the nature of the lesion and therefore, the prognosis.

Comparative Studies of the AgNORS Motifs in Phytohemagglutinin-Stimulated Human T-Lymphocytes with T-Lymphocyte Subgroups

OZET Objective: Using of argyrophilic nucleolar organiser region (AgNOR) patterns, instead of the fluorescent T-lymphocyte markers, in specification of sub-groups of T-lymphocyte was researched in the study. For this, it was aimed to indicate that AgNOR pattern of each T-lymphocyte sub-group remains stable in a stage of cell cycle stimulated by phytohaemagglutinin (PHA) and that these patterns are different from each other. Material and Methods: Peripheral blood samples of four healthy volunteers were cultured according to whole blood and/or isolated blood culture methods. The slides were stained by using AgNOR and immunohistochemical staining methods. Results: As we expected, the CD8 staining is not observed in a single style of the nucleolar organiser region (NOR) motif, but rather occurred in the forms of different kinds of nucleolus. It was found that the different types of NOR motifs formed with AgNOR staining are not related to the Tlymphocyte subgroup. Conclusion: It was concluded that the NOR motifs are not specific to sub-groups of T-lymphocytes. Further investigations are needed to support our results.

Patterns of nucleolar organizer region in urinary bladder cancer in view of 2004 WHO classification of urothelial neoplasms

Context: The last decade has seen a tremendous upheaval since introduction of WHO-1998 classification (modified in 2004) of bladder carcinoma, which only recently seems to have settled down. We compare both old (WHO-1973) and new (2004) classifications with argyrophilic nucleolar organizer region (AgNOR) score. Aims: To compare WHO/ISUP (World Health Organization/International Society of Urologic Pathologists) consensus classification-2004 and WHO-1973 classification of bladder carcinoma and to establish correlation between AgNOR scores and various grades of carcinoma bladder. Materials and Methods: Sixty H and E (Hematoxylin and Eosin) stained sections of transitional cell carcinoma bladder were graded according to WHO/ISUP (2004) and WHO (1973) classification systems and the results were compared. This was then followed by AgNOR staining of the paraffin-embedded sections and subsequent AgNOR scores were calculated and given as total score, extranucleolar score, intranucleolar score, and nucleolar score. These were correlated with the grade of the tumor. Statistical Analysis Used: ANOVA test. Results: It was observed that the two grading systems, WHO/ISUP (2004) and WHO (1973) correlated with each other. Also it was observed that the total AgNOR score and extranucleolar AgNOR score increased with the grade of the tumor and thus increasing scores carry significance in assessment of the grade of the tumor. However, intranucleolar and nucleolar scores showed variable degree of overlap in various grades of the bladder tumors. Conclusions: WHO/ISUP (2004) classification correlated well with the WHO (1973) classification. The total AgNOR score was found to be helpful in assessment of the grade of the tumor but the distribution of the AgNOR carried little significance.

Nucleolar organizer regions in a chronic stress and oral cancer model

This study aimed to examine the role of chronic restraint stress (RS) on oral squamous cell carcinomas induced by 4-nitroquinoline-1-oxide (4-NQO) in CF-1 mouse tongues, measured by the expression of argyrophilic staining of nucleolar organizer regions (AgNOR). Thirty one samples of lingual epithelial tissue of CF-1 mice with a diagnosis of oral squamous cell carcinoma (OSSC) were assigned to two experimental groups: the RS/4-NQO group, where animals received RS and induction of oral chemical carcinogenesis (n=17); and the 4-NQO group, where animals received induction of chemical carcinogenesis without restraint stress (n=14). The mean number and distribution pattern of AgNOR were recorded. The mean AgNOR number per cell was found to be slightly higher in the 4-NQO group. AgNOR in the RS/4-NQO group revealed a higher tendency to be arranged in a clumped distribution compared to the 4-NQO group. No statistically significant difference was found between the groups. In conclusion, the induction of chronic restraint stress in CF-1 mice does not increase the number or affect the distribution pattern of AgNORs in OSSC induced by 4-NQO.

Role of argyrophilic nucleolar organizer region staining in identification of malignant cells in effusion

Background:Cytological examination of effusions helps to differentiate between benign and malignant effusions, but fails a definitive diagnosis in a number of cases. The main problem here is to distinguish reactive mesothelial cells from neoplastic cells.Aim:In the present study, we used argyrophilic nucleolar organizer region (AgNOR) staining for diagnosis of benign and malignant effusions.Materials and Methods:In this study, 100 cases of effusion samples were taken and centrifuged. Four smears were prepared from sediment, one each for Leishman, hematoxylin and eosin (H and E) staining and two for AgNOR staining. AgNORs were counted as black dots in the nuclei of all abnormal cells using ×100 oil immersion objective. Their dispersion and shape was compared in benign, malignant and atypical cases.Results:Out of 50 pleural effusion samples, 29 were benign, 10 were malignant and 11 atypical. Among the 47 peritoneal effusion samples, 27 were benign, 16 were malignant and 4 atypical. Out of three cases of pericardial effusion samples, one was benign and two were malignant. All effusions were subjected to AgNOR staining. The benign group consisted of cells showing 1 to 2 dots of regular size and shape. In malignant group, 3 to 5 dots of variable size, shape and irregular contours were observed per cell. In atypical group, the reactive mesothelial cells showed 1 to 2 dots, malignant cells showed 3 to 4 irregular dots. Thus, a differentiation was noted between activated mesothelial cells and malignant cells, which was not possible in Leishman and H and E-stained smears alone.Conclusion:Therefore, AgNOR can be branded as an extremely useful additional diagnostic tool for cytodiagnosis of effusions.

Demonstration of disturbed activity of the lateral amygdaloid nucleus projection neurons in depressed patients by the AgNOR staining method

Background The aim to find a morphological biomarker of disturbed activity of the lateral amygdaloid nucleus in depression was approached by a karyometric analysis of projection neurons. Methods The study was performed on paraffin-embedded brains from 19 depressed patients from both the major depressive disorder (MDD) and the bipolar disorder (BD) diagnostic groups, including 10 suicides, and 24 matched controls. The karyometric parameters of the lateral amygdaloid nucleus (La) projection neurons bilaterally were evaluated by the argyrophilic nucleolar organiser region (AgNOR) silver staining method. Results An increased AgNOR number was found in the right La in suicides compared to controls. The intra-group comparisons between the hemispheres suggest a disturbed amygdaloid lateralisation in depressed patients. The effects were independent from psychotropic medication. There was a strong positive correlation between the nuclear area in La projection neurons and prefrontal limbic areas pyramidal neurons in the right hemisphere specific for suicide and MDD. Limitations A major limitation of this study is the relatively small number of cases. A further limitation is given by the lack of data on drug exposure across the entire lifespan. Conclusion The results suggest that depressed patients from both the MDD and BD diagnostic groups exhibit an increased activity of the La output neurons specific for suicidal patients. The distinctness of the diagnostic groups of mood disorders was accentuated in the correlation analysis. This putative hyperactivity was specific for the right hemisphere and psychotropic medication most likely did not counteract it.

Argyrophilic nucleolar organiser region (AgNOR) study of some odontogenic cyst epithelium

This study compared the biologic activity of three different cyst types and assessed the value of the AgNOR technique as an aid to diagnosis. The nucleolar organiser region (NORs) reflect replicatory activities within the cells. It has been found to be of diagnostic value in the prediction of the biologic behaviour of some tumors and even oral epithelial dysplasia. Thirty histologically diagnosed H & E sections of cystic lesions consisting of ten (10) radicular, ten (10 dentigerous and ten (10) odontogenic kerotocysts were randomly selected from the record of biopsy services of the Lagos University Teaching Hospital. The paraffin blocks of the selected sections were retrieved for AgNOR staining. For each cyst type the mean and standard deviation of AgNORs/100 cells was calculated and compared with other cyst types using an analysis of variance and student's t-tests. RESULTS of this study show a statistically significant higher means AgNORs cyst. (p = 0.0016; p = 0.00049 and respectively). However, no statistically significant difference in mean AgNOR score/cell was found when the radicular and dentigerous cysts were compared. These variations reflect the difference in the proliferation rate of the odontogenic keratocyst, radicular and dentigerous cysts.

Correlation Between Argyrophilic Nucleolar Organizer Region Staining and Brain Tumor Classification and Grading

ABSTRACTDistinctions between benign and malignant tumors are less evident in the central nervous system than in other tissues. Since the level of cell proliferation is an important feature in tumor grading, we applied AgNOR in 50 cases of brain tumors with different grades and histological origins in order to check its efficiency in discriminating between benign and malignant cases. We found significant differences between the means of grade I (1.76) and grade IV (2.46) tumors. No significant differences were observed considering the same grading with distinct histological types or age of patients, reinforcing the efficiency of AgNOR.

Cytogenetic analysis of orange-spotted grouper, Epinephelus coioides, using chromosome banding and fluorescence in situ hybridization

The orange-spotted grouper, Epinephelus coioides, is one of the most important commercial fish species in Southeast Asia. Although it has been widely cultured in China, basic cytogenetic information on this species is limited. In this study, we used Giemsa-staining, C-banding, argyrophilic nucleolar organizer regions (Ag-NORs), and repetitive sequencing (including 5S, 18S, and (TTAGGG)n) techniques to analyze the cytogenetics of the orange-spotted grouper. Our results showed that the karyotypic formula of the orange-spotted grouper was 2n = 2SM + 46A, NF = 50. C-banding patterns indicated that centromeric regions of all chromosomes were C-band positive except in pair no. 12. Heterochromatin was detected at the subcentromeric regions of both pair no. 5 and no. 12. In the smallest chromosome pair, no. 24, either the entire short arm or the end of the short arm was C-band positive. After Ag-NORs staining, one pair of nucleolar organizer regions was observed on the short arm of pair no. 24. FISH results showed that 5S rDNA was located at a pair of A-type chromosomes, while 18S rDNA appeared at the same location in the short arm of pair no. 24 where Ag-NORs were detected. The telomeric sequence (TTAGGG)n detected by FISH was located at both ends of each chromosome. Some of these chromosomal markers showed species-specific variations. Thus, despite the apparent karyotype stability (2n = 48), several microstructural chromosome changes must have occurred during chromosomal evolution, resulting in significant changes in the karyotype structure during the evolutionary diversification of the genus Epinephelus.