Argyrophilic Nucleolar Organizer Regions Research Articles

Computer-Assisted Morphometric Comparative Analysis of Argyrophilic Nucleolar Organizer Regions (AgNORs) in Leukoplakia With Dysplasia and Oral Squamous Cell Carcinoma.

Background Oral squamous cell carcinoma (OSCC) and oral leukoplakia (OL) with dysplasia are closely linked conditions in the oral cavity, with the latter often indicating precancerous changes, underscoring the urgency of early detection and intervention. Histopathological confirmation is crucial for accurate diagnosis. The nucleolar organizer region (NOR), specifically analyzed through silver-staining (argyrophilic NORs), provides insights into nuclear changes associated with the lesion. Computer-assisted morphometric analysis enhances precision and objectivity in evaluating AgNOR-related parameters. Aim To conduct a computer-assisted morphometric comparison of AgNORs using various NOR-related parameters in cases of OSCC and leukoplakia with dysplasia and to evaluate their diagnostic significance. Materials and methods A computer-assisted morphometric analysis was conducted using various NOR-related parameters, such as nuclear profile area, single AgNOR profile area per nucleus, total AgNOR profile area per nucleus, and number of AgNOR profiles per nucleus on a total sample of 90 specimens, which includes leukoplakia with dysplasia (30), OSCC (30), and a control group, including 30 samples of normal oral mucosa. A comparison was conducted on the morphometric values between the groups under investigation. Tukey's multiple comparison tests and ANOVA were used to analyze the data and determine the differences between the groups. Results The present investigation revealed a significant difference in all four AgNOR-related parameters between leukoplakia and OSCC in comparison to the control group (normal oral mucosa). Comparing OL (41.78 ± 0.46) and OSCC (62.78 ± 0.47) to the control group (35.93 ± 0.99), the mean value of nuclear profile area (A Nuc) was significantly greater. In comparison to the control group (3.40 ± 0.09), the mean value of a single AgNOR profile area per nucleus (A NOR) was found to be relatively lower in both research groups, OL (2.00 ± 0.02) and OSCC (1.39 ± 0.01). The total AgNOR profile area per nucleus (TA NOR) had a mean value of 10.61 ± 0.69 in OL and 12.05 ± 0.28 in OSCC, respectively, compared to 7.82 ± 0.38 in the control group. The study found that there was more number of profiles of AgNORs per nucleus (n NOR)in the study groups of OL (5.30 ± 0.29) and OSCC (8.69 ± 0.19) than in thecontrol group (2.32 ± 0.11). Conclusion The parameters linked to the NOR are biologically informative and easy to check regularly in a pathology lab. Additionally, AgNORs give us important information that enables us to study the range of nuclear changes in malignant and potentially malignant lesions.

Cutaneous Canine Mast Cell Tumor: The Use of Proliferative Markers (Ki-67 and Ki-67 × AgNOR) in Cytological Samples for Diagnosis and Prognosis.

A cytological grading system for canine mast cell tumors (MCTs) has been developed, but its integration into clinical routine has been hindered due to its diagnostic limitations. The aim of this study was to assess the prognostic value of Ki-67 and argyrophilic nucleolar organizing region (AgNOR) markers in cytological MCT samples and to determine cut-off values for these markers in correlation with histopathological grading. Cytological samples were collected prior to surgical excision, and histopathological samples were obtained postsurgery from 45 dogs diagnosed with cutaneous mast cell tumors (MCTs). The cytological specimens were classified using a two-tier grading system, and their Ki-67 (average immunopositive nuclei per 100 cells) and AgNOR (average AgNOR counts per 100 nuclei) signaling was assessed. Through receiver operating characteristic (ROC) analysis, cut-off values for Ki-67 and Ki-67 × AgNOR were determined to better align with histopathological grading (classified as low or high grade according to Kiupel's scoring system). Without the inclusion of proliferative markers, there was a 73% agreement between cytological and histopathological grading. The prediction of histopathological grade was slightly more accurate when assessing Ki-67 and Ki-67 × AgNOR signaling in cytological specimens (75% and 80%, respectively) compared to the initial cytological grading. The cytological assessment of canine MCTs proves beneficial for the initial evaluation, and the incorporation of the evaluation of Ki-67 and AgNOR markers may assist in identifying diagnostically highly malignant MCTs.

Investigation of the AgNOR (Argyrophilic Nucleolar Organizing Region) Protein Levels in Patients with Coronary Artery Diseases

Investigation of the AgNOR (Argyrophilic Nucleolar Organizing Region) Protein Levels in Patients with Coronary Artery Diseases

A CNN-based approach for joint segmentation and quantification of nuclei and NORs in AgNOR-stained images

Background and ObjectiveOral cancer is the sixth most common kind of human cancer. Brush cytology for counting Argyrophilic Nucleolar Organizer Regions (AgNORs) can help early mouth cancer detection, lowering patient mortality. However, the manual counting of AgNORs still in use today is time-consuming, labor-intensive, and error-prone. The goal of our work is to address these shortcomings by proposing a convolutional neural network (CNN) based method to automatically segment individual nuclei and AgNORs in microscope slide images and count the number of AgNORs within each nucleus. MethodsWe systematically defined, trained and tested 102 CNNs in the search for a high-performing solution. This included the evaluation of 51 network architectures combining 17 encoders with 3 decoders and 2 loss functions. These CNNs were trained and evaluated on a new AgNOR-stained image dataset of epithelial cells from oral mucosa containing 1,171 images from 48 patients, with ground truth annotated by specialists. The annotations were greatly facilitated by a semi-automatic procedure developed in our project. Overlapping nuclei, which tend to hide AgNORs, thus affecting their true count, were discarded using an automatic solution also developed in our project. Besides the evaluation on the test dataset, the robustness of the best performing model was evaluated against the results produced by a group of human experts on a second dataset. ResultsThe best performing CNN model on the test dataset consisted of a DenseNet-169 + LinkNet with Focal Loss (DenseNet-169 as encoder and LinkNet as decoder). It obtained a Dice score of 0.90 and intersection over union (IoU) of 0.84. The counting of nuclei and AgNORs achieved precision and recall of 0.94 and 0.90 for nuclei, and 0.82 and 0.74 for AgNORs, respectively. Our solution achieved a performance similar to human experts on a set of 291 images from 6 new patients, obtaining Intraclass Correlation Coefficient (ICC) of 0.91 for nuclei and 0.81 for AgNORs with 95% confidence intervals of [0.89, 0.93] and [0.77, 0.84], respectively, and p-values < 0.001, confirming its statistical significance. Our AgNOR-stained image dataset is the most diverse publicly available AgNOR-stained image dataset in terms of number of patients and the first for oral cells. ConclusionsCNN-based joint segmentation and quantification of nuclei and NORs in AgNOR-stained images achieves expert-like performance levels, while being orders of magnitude faster than the later. Our solution demonstrated this by showing strong agreement with the results produced by a group of specialists, highlighting its potential to accelerate diagnostic workflows. Our trained model, code, and dataset are available and can stimulate new research in early oral cancer detection.

Association of Proliferative Indices With Various Grades of Breast Carcinoma.

Various grades of breast carcinoma and proliferative indices used as nuclear protein Ki-67 and argyrophilic nucleolar organizer regions (AgNOR) are being compared to each other. In this observational cross-sectional investigation, 42 breast biopsies from questionable breast areas were collected and preserved in formalin and paraffin before the tissue blocks were made. A thorough medical history regarding the breast tumor and thorough physical examination results were recorded. Two sections were produced, one stained with an immunohistochemical marker called Ki-67 and the other with a unique stain called AgNOR. Grade I in Nottingham was found to be highest in subjects with Ki-67 1%, grade II in subjects with Ki-67 1-10%, and grade III in subjects with Ki-67>10%. Therefore, a higher Ki-67 score and a higher Nottingham grade were more closely associated. The mean AgNOR score was determined to be highest in Nottingham grade III and lowest in Nottingham grade I. In contrast to grade I and grade II of carcinoma (CA) breast, where there was no statistically significant association between Ki-67 and AgNOR, grade III of CA breast showed a statistically significant link between Ki-67 and AgNOR. Proliferation has been identified as a distinctive feature of cancer and as a key factor in the prognosis of the disease.

Chemopreventive Effect of Cooked Chickpea on Colon Carcinogenesis Evolution in AOM/DSS-Induced Balb/c Mice.

Chickpeas are one of the most widely consumed legumes worldwide and they might prevent diseases such as cancer. Therefore, this study evaluates the chemopreventive effect of chickpea (Cicer arietinum L.) on the evolution of colon carcinogenesis induced with azoxymethane (AOM) and dextran sodium sulfate (DSS) in a mice model at 1, 7, and 14 weeks after induction. Accordingly, the expression of biomarkers-such as argyrophilic nucleolar organizing regions (AgNOR), cell proliferation nuclear antigen (PCNA), β-catenin, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2)-was assessed in the colon of BALB/c mice fed diets supplemented with 10 and 20% cooked chickpea (CC). The results showed that a 20% CC diet significantly reduced tumors and biomarkers of proliferation and inflammation in AOM/DSS-induced colon cancer mice. Moreover, body weight loss decreased and the disease activity index (DAI) was lower than the positive control. Lastly, tumor reduction was more evident at week 7 in the groups fed a 20% CC diet. In conclusion, both diets (10% and 20% CC) exert a chemopreventive effect.

Evaluation of the Cytological Changes of the Oral Mucosa Among Smokers in Al Madinah Al Munawara Using Argyrophilic Nucleolar Organizer Region (AgNOR) Counts and Papanicolaus Stain.

Objective To evaluate the cytological changes of the oral mucosa among smokers using Argyrophilic nucleolar organizer region (AgNOR) counts and Papanicolaou (Pap) staining. Methodology The oral mucosal exfoliate smears of 500 individuals (200 nonsmokers and 300 smokers) aged between 18 and 80 years were prepared in Al Madinah. TheAgNOR count and Pap stain were used to generate a cytogenic smear to assess the presence of cytological changes suggestive of atypia, inflammation, dysplasia, keratinization, and proliferative activity of epithelial cells. Results Smokers have a considerably higher number of AgNORs per nucleus than nonsmokers (1.99 3.53 vs. 0.42 1.22). There were inflammatory changes in 127 (42.3%) of the cases and 40 (20%) of the controls. Multinucleated cells and atypia were found in 33 (11%) and 14 (4.5%) of the casesbut not in the controls. The results indicate higher proliferative activity in smoking patients compared to nonsmoking patients, even in the absence of clinical lesions. Conclusion To detect the effects of smoking on the oral mucosa, Pap staining alone is insufficient. Combining Pap staining with the AgNOR technique produces the desired results.

MDA-MB-231 Human Breast Cancer Cell Line Treated with Ginseng (Panax Quinquefolius): Evaluation by Annexin V and AgNOR Staining

Aim: In this study, it was aimed to examine the time and dose dependent effects of Ginseng (Panax quinquefolius) on MDA-MB-231 cell lines. Material and Methods: MDA-MB-231 breast cancer cell line was used in the study. MDA-MB-231 cells were exposed to ginseng at 37°C and 5% CO2 for varying durations (24 and 48 hours) and doses (1 and 2 μg/ml ginseng). At the end of the incubation period, viability, apoptosis, cell cycle and Argyrophilic nucleolar organizing region (AgNOR) protein status of MDA-MB-231 cells were examined in the Muse Cell Analyzer. Results: It was observed that the dose inducing apoptosis was 1 μg/ml ginseng for 24 and 48 hours, and 2 μg/ml ginseng for 48 hours in the group that stopped the cell cycle in the G0/G1 phase. When comparing the two groups; while no difference was determined between the control and 1 µg/ml ginseng groups, the significant differences were detected between the control and 2 µg/ml ginseng groups for mean AgNOR number in 48 hours incubation. However, there was no significant difference for the TAA/NA ratio, in the groups for 48 hours. Conclusion: The current study showed that ginseng had a crucial function against cancer development. Also, both AgNOR values might be used as biomarkers for detection of the most reliable therapeutic dose selection for cancer and it has been shown that correct consumption of Ginseng can be effective in preventing cancer formation and slowing its progression.

Argyrophilic nucleolar organizer regions as a promising biomarker for the detection of brain hypoxia levels caused by different doses of carbon monoxide poisoning.

The purpose of the study is to investigate whether there is any relationship between mean argyrophilic nucleolar organizing regions (AgNOR) number and total AgNOR area/total nuclear area (TAA/TNA) ratio and the levels of brain hypoxia after exposure to different acute doses of carbon monoxide (CO) gas. Each experimental group was exposed to CO gas (concentrations of 1,000, 3,000 and 5,000 ppm). Then, the rats were anesthetized, and blood samples were taken from the right jugular vein for carboxyhemoglobin levels detection. The rats were sacrificed on seventh day. AgNOR staining was applied to brain tissues. TAA/TNA and mean AgNOR number were detected for each nucleus. Significant differences were detected among all groups for TAA/TNA ratio, mean AgNOR number and carboxyhemoglobin level. According to a double comparison of groups, the differences between control and 1,000 ppm, control and 3,000 ppm, control and 5,000 ppm, and between 1,000 and 5,000 ppm were significant for TAA/TNA ratio. When mean AgNOR number was considered, significant differences were detected between control and 1,000 ppm, control and 3,000 ppm, control and 5,000 ppm, and between 1,000 and 3,000 ppm. AgNOR proteins may be used for early detection of the duration, intensity, and damage of brain injury caused by CO poisoning. Thus, effective treatment strategies can be developed for the prevention of hypoxic conditions.

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.

Use of AgNOR staining to determine the effect of metoclopramide on neural tube development in early chick embryos

ABSTRACT Nausea and vomiting during pregnancy are common problems and prolonged pharmacological treatment often is needed; however, the teratogenic effects of anti-emetic drugs on neural tube (NT) development are not clear. We investigated the effects of different doses of metoclopramide on NT development in 48 and 72 h chick embryos using an argyrophilic nucleolar organizing region (AgNOR) staining method. We used 150 fertile, specific pathogen-free eggs incubated for 28 h, then randomly divided into five equal groups: group A, sham control was administered 0.9% saline; groups B – E were administered 0.15 mg/egg, 0.3 mg/egg, 0.6 mg/egg and 1.2 mg/egg, respectively. Half of the eggs in each group were taken from the incubator at 48 h incubation and the other half at 72 h incubation. After incubation, eggs were opened, embryos were dissected from their membranes, fixed with 10% formalin and examined by light microscopy. The NT status, i.e., open or closed, and somite number, crown-rump length, morphological features and gross developmental abnormalities were recorded. Excised embryos were sectioned and stained using hematoxylin and eosin or the AgNOR procedure and examined for morphology and histopathology. Delayed NT closure was observed in all 48 h drug exposed embryos, but in the 72 h groups, this occurred only in high-dose groups. Somite number was reduced significantly in groups C – E compared to the control group. Crown-rump length was decreased in both 48 and 72 h embryos. We found a decreased total AgNOR area:nuclear area ratio in 48 and 72 h embryos of all experimental groups. We found that metoclopramide delayed NT closure in chick embryos in a dose-dependent manner.

A Comparison of Proliferative Capacity of Reticular and Erosive Variants of Oral Lichen Planus by Argyrophilic Nucleolar Organizer Regions Method.

Oral lichen planus (OLP), an immune mediated disorder, has been recognized since 1869 and is presented as any one of the six variants. Reticular and erosive are the most frequently encountered. Its proliferative capacity can give some information regarding its progression. We adopted the argyrophilic nucleolar organizer regions (AgNORs) method because of its simplicity to use and dependable results. We evaluated AgNORs in basal, suprabasal, and squamous cell layers. We also compared these three layers within two variants, reticular, and erosive. Thirty clinically diagnosed patients of OLP were included in the study. Reticular and erosive variants were included in our study. This was followed by hematoxylin and eosin staining and later by the AgNOR method. The mean number of AgNORs per nucleus was calculated. Thirteen males and 17 females were the gender distribution. Twenty-three (76.67%) had reticular pattern and seven (23.33%) had erosive pattern. The basal cell layer had the highest mean AgNOR compared to suprabasal and squamous layers. Even among, erosive and reticular variants, the former had higher mean AgNOR counts. Our results suggest that the inflammatory infiltrate close to the epithelial cells can alter the proliferation index for the pattern of protein synthesis of these cells. Moreover, the high proliferative index in OLP can be related to a specific immunologic response. We conclude that AgNOR can be used as a proliferative marker in earlier lesions to detect the severity.

Relationship Between Argyrophilic Nucleolar Organizer Region Proteins of Pediatric Epilepsy and T Lymphocyte Subpopulation

Relationship Between Argyrophilic Nucleolar Organizer Region Proteins of Pediatric Epilepsy and T Lymphocyte Subpopulation

Prognostic Potential of Argyrophillic Nucleolar Organizer Regions (Agnors) in Oral Lesions: A Systematic Review

Early detection of oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMD) is important for dental professionals to improve patient survival rates. More than half of patients with oral squamous cell cancer had advanced disease at the time of diagnosis, indicating a lack of early detection and risk assessment biomarkers. The development of new protein biomarkers will help in early diagnosis and treatment.The argyrophilic nucleolar organiser regions (AgNORs) staining technique is simple and cost-effective. These are replicatory markers that identify epithelial dysplasia. And it also plays a very important role in differentiating the benign, pre-malignant, and malignant lesions of the oral cavity. The number of AgNORs per cell has been considered an indicator of cellular proliferative activity. Microscopically, NORs can be identified as well-defined black dots located throughout the cell nucleus. The agNOR quantity is strictly proportional to the proliferative activity of the cell. AgNOR quantification helps in the determination of the degree of epithelial dysplasia and, consequently, in the analysis of its potential for malignant transformation. AgNOR qualitative characteristics help in differentiating hyperplastic, premalignant, and malignant oral lesions. The sliver staining technique is useful for studying the structure of the nucleolus as well as the variations in its activity. AgNORs are a valuable parameter in tumour pathology.

A Study of AgNOR Scores on FNAC of Breast Lumps and Its Correlation with Histopathological Grading

Background: Breast cancer is one of the leading cancers in women both in developed and developing countries. Nucleolar Organizer Regions (NORs) are loops of DNA related to protein synthesis and ribosomal activity. Argyrophilic Nucleolar Organizer Region (AgNOR) counts act as a reliable cell proliferative marker as they aid in identifying cell proliferation activity and aggressiveness of a lesion, and thereby helps in differentiating benign and malignant breast lesions.&#x0D; Aim: To evaluate the role of AgNOR count in differentiating benign and malignant breast lumps.&#x0D; Methods: All breast lump specimens collected for Fine Needle Aspiration Cytology (FNAC) and histopathology, for a period of one year, were included in the study. A total of 69 specimens were included. All FNAC and histopathology slides were stained with AgNOR stain, and the score was recorded.&#x0D; Result: A total of 69 cases were studied, both non-neoplastic and neoplastic breast lesions. All FNAC cases correlated histologically. Out of the 69 cases, 38 were malignant. The mean AgNOR counts in non-neoplastic, benign and malignant breast lesions were found to be 2.19, 2.88 and 5.94 respectively (p&lt;0.05). Other parameters such as proliferative AgNOR index, AgNOR size variation, AgNOR distribution within the nucleus and the Subjective AgNOR Pattern Assessment (SAPA) scores also showed statistically significant difference between benign and malignant lesions.&#x0D; Conclusion: AgNOR scores showed good correlation in differentiating benign and malignant breast lesions, they can be used as a reliable tool to aid in the diagnosis.

Could Argyrophilic Nucleolar Organizer Regions count mirror DNA Ploidy in Malignant Salivary Gland Tumors?

Objective:Nucleolar organizer regions (NORs) are DNA coils that transcribe to ribosomal RNA. The NOR-associated protein, termed argyrophilic NOR (AgNOR), was visible within the nucleus by staining with silver nitrate examination via the light microscope. AgNOR counting is a proliferation marker and may help in the diagnosis and prognosis of various neoplastic lesions. Aneuploidy (abnormal DNA content) can predict the progression, survival and prognosis of the tumors. The aim of this study was to evaluate the role of AgNORs, DNA ploidy status, and total S-phase fraction (TSPF) as prognostic parameters in malignant salivary gland tumors (MSGTs). Methods:The current study is a retrospective study on a cohort of MSGTs (N=47), to assess AgNORs using Silver Nitrate stain, DNA index (DI), and TSPF using flow cytometry (FCM). Data including tumor size and site, lymphovascular invasion (LVI), lymph node metastasis (LNM) were collected. Results:The AgNORs count was statistically significant with MSGT type. DI was found to have a significant association with tumor site, tumor size and MSGT type. In addition, TSPF was found to be significantly associated with LVI. A moderate positive correlation was noted between AgNORs count and TSPF. LNM, tumor site, high AgNORs and low DI were all associated with short disease-free survival (DFS) and poor overall survival (OS). Conclusion:The present study revealed that high AgNORs count, DNA aneuploidy and TSPF had a poor influence on MSGTs prognosis.

Ribosomal DNA transcription is increased in the left nucleus accumbens of heroin-dependent males.

Opioid addiction is a worldwide problem accentuated in the USA and European countries by the COVID-19 pandemic. The nucleus accumbens (NAc) plays an outstanding neurobiological role in opioid addiction as a part of the striatum and key component of brain reward system. The striatal GABAergic medium spiny projection neurons (MSNs) are the main neuronal type in the NAc where addiction-specific synaptic plasticity occurs. The activity of ribosomal DNA (rDNA) transcription is crucial for neural plasticity and molecular studies suggest its increase in the NAc of heroin addicts. Silver-stained argyrophilic nucleolar organizer region (AgNOR) areas visualised in neuronal nuclei in paraffin-embedded brain sections are reliable morphological estimators of rDNA transcription and thus surrogate markers for the activity of brain regions. Our study revealed increased AgNOR areas in MSNs of the left NAc in 11 heroin addicts versus 11 healthy controls from the Magdeburg Brain Bank (U-test P = 0.007). No differences were observed in another investigated part of the striatum, namely the head of caudate nucleus, which is located closely to the NAc. The results were not confounded by significant differences in the age, brain volume and time of formalin fixation existing between compared groups. Our findings suggest an increased NAc activity in heroin addicts, which is consistent with human and animal experimental data.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00406-022-01423-7.

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.

Τhe Effect of Opioid Administration on Cytologic and Histopathologic Diagnosis of Canine Cutaneous Mast Cell Tumors Treated by Surgical Excision.

Mast cell tumor (MCT) is a frequent cutaneous tumor in dogs, with a variable biological behavior. Studies correlate cytologic and histopathologic features of MCTs with their biological behavior, prognosis, and response to treatment. The use of preoperative opioids is common in canine patients undergoing surgical removal of these tumors. Certain opioids can induce or downregulate mast cell degranulation and influence cancer progression. The aim of the present study was to investigate whether the administration of morphine or butorphanol during surgical excision of canine cutaneous MCTs affects their cytologic and histopathologic appearance, thus influencing cytologic and histopathologic grading. This was a prospective, blinded, randomized, cohort clinical study. Forty-five dogs with cutaneous MCTs were randomly allocated into three groups according to preanaesthetic medication: dexmedetomidine combined with morphine (group M) or butorphanol (group B) or normal saline (group C). Cytologic specimens and histopathologic samples were obtained both prior to and after surgery. Samples were graded according to Kiupel’s and Patnaik’s systems, examined immunohistochemically for Ki-67 protein (Ki-67) and c-kit proto-oncogene product (KIT) expression, and histochemically for argyrophilic nucleolar organizing regions (AgNORs). Based on both Kiupel’s and Patnaik’s systems, no statistically significant differences were noted concerning the number of cases with grading discrepancies in grades allocated prior to versus after surgery among the groups. The same applied for cytological grading and immunohistochemical and histochemical evaluation. It seems that administration of morphine or butorphanol as part of the preanesthetic medication for surgical removal of canine cutaneous mast cell tumors does not influence histopathologic and cytologic grading of MCTs.

“STUDY OF PROLIFERATIVE INDICES USING MITOTIC INDEX AND AGNOR AND THEIR CORRELATION WITH VARIOUS GRADES OF BREAST CARCINOMA”

Background: Breast cancer is a leading cause of death in women. The objectives of this study were to compare the argyrophilic nucleolar organizer region (AgNOR) count and mitotic index with various histological grades of infiltrating ductal carcinoma of breast. Aim: To study the relationship of the proliferative indices and various grades of breast carcinoma. Objective: "Study of proliferative indices using mitotic index and Agnor and their correlation with various grades of breast carcinoma" Material and Method: In this prospective study all specimens of infiltrating ductal carcinoma of female breast belonging to all age groups received in the department of histopathology of Gajra Raja Medical College,Gwalior during the period of three years were graded using Bloom and Richardson score and correlated with proliferative indices mitotic index and AgNOR counts. Result: The mean Age (Years) was 46.16 ± 8.68.The mean B&amp;R Score was 6.52 ± 1.20.The mean Mitotic Index was 8.51 ± 2.61.The mean AgNOR Count was 4.24 ± 2.34.There was significant correlation between AgNOR count,mitotic index and histological grades. Conclusion: AgNOR count and mitotic index can be used as reliable proliferative indices which correlate well with the histological grades and are cost effective too.