Nucleolar Characteristics Research Articles

Tumor diagnosis based on nucleolus labeling

The relationship between the nucleolus and tumorigenesis was explored, and the current methods for diagnosing tumors were evaluated by examining nucleolar characteristics.

Cytogenotoxicity Screening of Urban and Rural Marshes: An Integrated In Vivo Approach Coupling Fish and Plant-Based Tests Adapted for Low-Income Countries.

Effects of anthropogenic activities such as urbanization, population growth, and agriculture on water quality are major concerns particularly in low-income countries where water quality monitoring can be challenging. The purpose of the present study was to evaluate the cytogenotoxic potential of water from urban and rural Malagasy marshes, coupling a fish (Nile tilapia, Oreochromis niloticus) and a plant (Allium cepa) species as bioindicators. The fish and plants were exposed for 72 h to water sampled in the two locations investigated. Using the comet assay on fish erythrocytes, DNA strand breaks were assessed, while mitotic index and nucleolar alterations were estimated in cells of the plant root apex. Comet assays revealed significant DNA strand breaks to fish erythrocytes in both the marshes investigated while the mitotic index and nucleolar characteristics in the roots of A. cepa mainly highlighted potential cytotoxicity in the urban marsh. Our results demonstrate the advantages of coupling in vivo biological test systems to screen potential cytogenotoxicity of surface water in low-income countries where comprehensive data sets of aquatic contaminants are often lacking. Environ Toxicol Chem 2023;42:1266-1275. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Nucleoli and the nucleoli-centromere association are dynamic during normal development and in cancer.

Centromeres are known to cluster around nucleoli in Drosophila and mammalian cells, but the significance of the nucleoli-centromere interaction remains underexplored. To determine whether the interaction is dynamic under different physiological and pathological conditions, we examined nucleolar structure and centromeres at various differentiation stages using cell culture models and the results showed dynamic changes in nucleolar characteristics and nucleoli-centromere interactions through differentiation and in cancer cells. Embryonic stem cells usually have a single large nucleolus, which is clustered with a high percentage of centromeres. As cells differentiate into intermediate states, the nucleolar number increases and the centromere association decreases. In terminally differentiated cells, including myotubes, neurons, and keratinocytes, the number of nucleoli and their association with centromeres are at the lowest. Cancer cells demonstrate the pattern of nucleoli number and nucleoli-centromere association that is akin to proliferative cell types, suggesting that nucleolar reorganization and changes in nucleoli-centromere interactions may play a role in facilitating malignant transformation. This idea is supported in a case of pediatric rhabdomyosarcoma, in which induced differentiation reduces the nucleolar number and centromere association. These findings suggest active roles of nucleolar structure in centromere function and genome organization critical for cellular function in both normal development and cancer.

Krüppel-like factor 7 influences translation and pathways involved in ribosomal biogenesis in breast cancer

BackgroundRibosomal biogenesis and ribosomal proteins have attracted attention in the context of tumor biology in recent years. Instead of being mere translational machineries, ribosomes might play an active role in tumor initiation and progression. Despite its importance, regulation of ribosomal biogenesis is still not completely understood.MethodsUsing Gene Set Enrichment Analysis of RNA sequencing and proteomical mass spectrometry data in breast cancer cells expressing Krüppel-like factor 7 (KLF7), we identified processes altered by this transcription factor. In silico analyses of a cohort of breast cancer patients in The Cancer Genome Atlas confirmed our finding. We further verified the role of KLF7 the identified ribosomal processes in in vitro assays of mammary carcinoma cell lines and analyses of breast cancer patients’ tissue slices.ResultsWe identified the transcription factor Krüppel-like factor 7 (KLF7) as a regulator of ribosomal biogenesis and translation in breast cancer cells and tissue. Highly significant overlapping processes related to ribosomal biogenesis were identified in proteomics and transcriptomics data and confirmed in patients’ breast cancer RNA Seq data. Further, nucleoli, the sites of ribosomal biogenesis, were morphologically altered and quantitatively increased in KLF7-expressing cells. Pre-rRNA processing was identified as one potential process affected by KLF7. In addition, an increase in global translation independent from proliferation and transcription was observed upon exogenous KLF7 expression in vitro. Importantly, in a cohort of breast cancer patients, KLF7-expression levels correlated with aggressiveness of the intrinsic breast cancer subtype and tumor grading. Moreover, KLF7 correlated with nucleolar characteristics in human breast tumor tissue, indicating a role for KLF7 in ribosomal biogenesis.ConclusionIn mammary carcinoma, KLF7 is involved in ribosomal biogenesis. Alterations of ribosomal biogenesis has far reaching quantitative and qualitative implications for the proteome of the cancer cells. This might influence the aggressiveness of cancer cells.

Nuclear Morphological Characteristics in Breast Cancer: Correlation with Hormone Receptor and Human Epidermal Growth Factor Receptor 2.

Background Hormone receptor (HR) and human epidermal growth factor receptor 2 (HER2) are the common diagnostic/prognostic markers in breast cancer. Few articles have recently reported the correlation between cytology and molecular subtypes. We combined nuclear morphological characteristics with HR and HER2 status to observe the relationship and provide ideas for machine learning. Methods We reanalyzed fine-needle aspiration cytology samples and core-needle puncture histological specimens from 142 patients with invasive breast cancer between March 2019 and December 2019, and the findings were compared with the two groups (HR+/HER2- and HR-/HER2+) following nuclear cytomorphological features: nuclear/cytoplasmic ratio, difference of nuclear size, nuclear pleomorphism, chromatin feature, nuclear membrane and nucleoli, and Nottingham grading. Results Two groups were significantly associated with the difference of nuclear size, nuclear pleomorphism, and nucleoli (P < 0.001) and consistent with histological grading (P < 0.001). Moreover, nucleolar characteristics of size and number had obviously statistical significance (P < 0.001). Multiple micro-nucleoli were frequently seen in the HR+/HER2- group compared with the HR-/HER2+ group which mostly were observed centered medium-large nucleoli. We described four interesting nuclear morphologies in the experiment. Conclusions There were significant differences in nuclear characteristics between two groups. HR and HER2 status not only might be predicted in cytological samples, but some specific nuclear morphological features might have potential value to help us understand molecular function and predict more information.

Rapid ex vivo assessment of cancer prognosis by fluorescence imaging of nucleolus using nitrogen doped carbon dots

Cancer severely threatens human health currently, promoting the rapid development of cancer treatment strategies. In addition to cancer therapy, assessment of cancer prognosis, which can evaluate the success with treatment and chances of recovery as well as assist to make subsequent therapeutic schedule, is also remarkably indispensable and important. Conventional technologies can’t provide rapid and highly-sensitive assessment of cancer prognosis at cytological level. Herein, an effective nitrogen doped carbon dots with intrinsic nucleolus-targeting capability and high fluorescence quantum yield are synthesized, characterized and employed for fluorescence imaging of nucleolus, which is closely related to the biological alteration of cancer cell. The cancer prognosis thus can be accurately (limit of detection: 50 nM) and rapidly (5 min) assessed at subcellular organelle level from nucleolar characteristics, which are visualized and analyzed by the captured fluorescence images. Outstanding assessment performance endows the proposed technology with great potential for future clinical research.

Nucleoli cytomorphology in cutaneous melanoma cells \u2013 a new prognostic approach to an old concept

BackgroundThe nucleolus is an organelle that is an ultrastructural element of the cell nucleus observed in H&E staining as a roundish body stained with eosin due to its high protein content. Changes in the nucleoli cytomorphology were one of the first histopathological characteristics of malignant tumors. The aim of this study was to assess the relationship between the cytomorphological characteristics of nucleoli and detailed clinicopathological parameters of melanoma patients. Moreover, we analyzed the correlation between cytomorphological parameters of nucleoli and immunoreactivity of selected proteins responsible for, among others, regulation of epithelial-mesenchymal transition (SPARC, N-cadherin), cell adhesion and motility (ALCAM, ADAM-10), mitotic divisions (PLK1), cellular survival (FOXP1) and the functioning of Golgi apparatus (GOLPH3, GP73).MethodsThree characteristics of nucleoli – presence, size and number – of cancer cells were assessed in H&E-stained slides of 96 formalin-fixed paraffin-embedded primary cutaneous melanoma tissue specimens. The results were correlated with classical clinicopathological features and patient survival. Immunohistochemical analysis of the above mentioned proteins was described in details in previous studies.ResultsHigher prevalence and size of nucleoli were associated with thicker and mitogenic tumors. All three nucleolar characteristics were related to the presence of ulceration. Moreover, microsatellitosis was strongly correlated with the presence of macronucleoli and polynucleolization (presence of two or more nucleoli). Lack of immunologic response manifested as no TILs in primary tumor was associated with high prevalence of melanoma cells with distinct nucleoli. Interestingly, in nodular melanoma a higher percentage of melanoma cells with prominent nucleoli was observed. In Kaplan-Meier analysis, increased prevalence and amount, but not size of nucleoli, were connected with shorter cancer-specific and disease-free survival.Conclusion(1) High representation of cancer cells with distinct nucleoli, greater size and number of nucleoli per cell are characteristics of aggressive phenotype of melanoma; (2) higher prevalence and size of nucleoli are potential measures of cell kinetics that are strictly correlated with high mitotic rate; and (3) high prevalence of cancer cells with distinct nucleoli and presence of melanocytes with multiple nucleoli are features associated with unfavorable prognosis in patients with cutaneous melanoma.

Pronuclear morphology evaluation in in vitro fertilization (IVF) / intracytoplasmic sperm injection (ICSI) cycles: a retrospective clinical review

BackgroundThe assessment of the embryo quality is crucial to maintain an high pregnancy rate and to reduce the risk of multiple pregnancy. The evaluation of the pronuclear and nucleolar characteristics of human zygote have been proposed as an indicator of embryo development and chromosomal complement. The aim of the current study was to assess the role of pronuclear morphology evaluation in vitro fertilization (IVF) / intracytoplasmic sperm injection (ICSI) cycles.MethodsRetrospective clinical analysis on 755 non-elective transfers of only one embryo (ET). Embryo assessment was performed in days 1 and 2. Clinical and biological data were recorded and analyzed according to embryo and/or pronuclear morphology.ResultsBoth pronuclear and embryo morphology were significantly related to clinical pregnancy and live-birth rates. No significant difference in clinical pregnancy and live-birth rates was detected when the pronuclear and embryo morphology assessments were combined. Embryo morphology and maternal age were the only independent predictors of favorable outcome by logistic regression analysis.ConclusionsPronuclear evaluation is effective to select the best zygotes if ET is performed at day 1, whereas it did not improve the clinical outcomes when combined with embryo morphology evaluation in day 2.

Long-term toxicity of hexabromocyclododecane (HBCDD) to the benthic clam Macoma balthica (L.) from the Baltic Sea

The world's largest brackish water sea area, the Baltic Sea, is considered to be one of the most polluted seas of the world. Many new pollutants are constantly entering the environment, such as brominated flame-retardants (BFRs). BFRs represent a group of compounds that structurally resemble hydrophobic organic contaminants, but only scarce data about their toxicity to marine organism exist. Thus, the purpose of this study was to analyze long-term in vivo cytotoxicity and genotoxicity of hexabromocyclodododecane (HBCDD) to exposed marine invertebrates using a suite of cytogenetic biomarkers. This included a set of nuclear and nucleolar characteristics and the micronucleus test. The use of those parameters reflects different mechanisms of nuclear activity regulation in cells (as a parameter of cytotoxicity) and measures subcellular processes. The induction of nuclear abnormalities (like the formation of micronuclei) was also employed here as a parameter of genotoxicity. In order to reflect the proliferative and metabolic activity of the cells the number of argylophillic nucleolar organiser regions (NORs) in interphase cells was scored. Over a period of 50 days an in vivo exposure experiment with a clam Macoma balthica and different concentrations of HBCDD (nominal concentrations of 0, 100 and 250 μg/l) with three replicates each was performed. Gill cells were used as “sentinel systems” considering specificity in metabolism, repair mechanisms, adaptative response and cell proliferation. A significant increase in nuclear and nucleolar abnormalities and in the frequency of dead cells was observed during the duration of the experiment with the highest peak occurring 10 days after exposure for nuclear abnormalities and 20–30 days after exposure for malfunction of ribosomal genes (NORs) (GLM analyses and Spearman correlation, p < 0.05). Thus, the induction of micronuclei and other nuclear abnormalities reflected the toxic potential of HBCDD to marine invertebrates while an increase in the number of NOR may also reflect adaptive responses of the system as enhanced induction of proliferative regeneration of the gill tissue.

Nucleolar variation in response to nutritional condition in juvenile pejerrey Odontesthes bonariensis (Valenciennes)

The variations of the nucleolar characteristics size and number were studied in the gill and liver of Odontesthes bonariensis under different feeding regimes in order to test the hypothesis that functional genomes at the nucleolus, which determine variation of size (heterogeneity) in juvenile fishes, are modulated by environmental factors such as food availability. The programme that controls genetic changes of the nucleolar characteristics of the diploid cells of O. bonariensis was different from that described in the diploid cells of cyprinid fishes. No variations of the mean nucleolar number per nucleus were observed in liver or gill cells in relation to the feeding regime. Conversely, significant reduction of the mean nucleolar volume per nucleus (VMN) and change in the shape of the frequency distribution of the total nucleolar volume per cell was evident in both cell types in response to starvation or food restriction. Tissue specificity was observed in relation to the absolute values of the nucleolar characteristics; however, the relative behaviour was comparable in digestive and non‐digestive tissues. The time‐course reduction of VMN in gill and liver cells of restrictively fed fish followed a negative exponential. A good correlation between VMN and the condition index of the fish (K) was observed after long‐term food restriction; however, a faster response of VMN was observed after short‐term fasting events. Results supported the stated hypothesis and showed nucleolar activity as a direct, rapid and sensitive indicator of the nutritional status of the fish.

Using the nucleolar biomarker and the micronucleus test on in vivo fish fin cells

This study was aimed at developing the nucleolar biomarker and the micronucleus test on in vivo fish fin cells for assessing water cytotoxicity and genotoxicity. Both biomarkers can be used either jointly or separately on fins of the same fish during the experiment. For studying the nucleolar characteristics, small pieces of the fin edge were cut several times during 30 – 180 min of fish exposure. For micronucleus testing, the fin tissue regenerating after its cutting was investigated after 2–5 days of fish incubation. Effects of copper (0.1 and 2.5 mg/L), cadmium (0.005 and 1.0 mg/L) ions and chloral hydrate (400 and 800 mg/L) solutions were studied on cells of common carp ( Cyprinus carpio L.), crucian carp ( Carassius auratus gibelio Bloch.), and Mozambique tilapia ( Tilapia ( Sautherodon) mossambica) using a set of nucleolar characteristics (the number of nucleoli per cell, the size of a single nucleolus, and the percentage of cells with heteromorphic paired nucleoli) and the frequencies of cells with micronuclei and double nuclei. Substantial changes in parameters of nucleolar activity of fin cells were found to be caused by cadmium and copper impact. In comparison to blood cells, gill and fin cells were more sensitive as demonstrated by their nuclear damages after the chloral hydrate influence. Fin cells were useful to determine periodically cytotoxic and genotoxic effects of organic and inorganic substances in the same individual fish without any disruption of its physiological functions.

Characterization of mouse spermatogonia by transmission electron microscopy.

Characteristics of the various type A, intermediate (In) and B spermatogonia were determined in C57BL/6J mice using transmission electron microscopy. Spermatogonia were photographed at all stages of the cycle of the seminiferous epithelium. Over 450 images were taken. Spermatogonia could be classified into As, Apr, Aal, A1 cells, A2 cells, A3 cells, A4 cells, intermediate type and type B cells primarily on the basis of nuclear and nucleolar characteristics. The most primitive spermatogonia (As, Apr, Aal) had mottled chromatin; A1 cells contained homogeneously finely granular chromatin throughout the nucleus; A2, A3, A4 and intermediate type spermatogonia had progressively increasing amounts of chromatin encrusting the nuclear envelope; type B spermatogonia had less heterochromatin along the nuclear envelope, although the particles were more dense and rounded than in intermediate type spermatogonia. Mitochondrial size and position of Golgi complexes varied in different types of spermatogonia. These data show that types of spermatogonia can be differentiated such that these characteristics can be used in functional studies.

Characterization of mouse spermatogonia by transmission electron microscopy

Characteristics of the various type A, intermediate (In) and B spermatogonia were determined in C57BL/6J mice using transmission electron microscopy. Spermatogonia were photographed at all stages of the cycle of the seminiferous epithelium. Over 450 images were taken. Spermatogonia could be classified into As, Apr, Aal, A1 cells, A2 cells, A3 cells, A4 cells, intermediate type and type B cells primarily on the basis of nuclear and nucleolar characteristics. The most primitive spermatogonia (As, Apr, Aal) had mottled chromatin; A1 cells contained homogeneously finely granular chromatin throughout the nucleus; A2, A3, A4 and intermediate type spermatogonia had progressively increasing amounts of chromatin encrusting the nuclear envelope; type B spermatogonia had less heterochromatin along the nuclear envelope, although the particles were more dense and rounded than in intermediate type spermatogonia. Mitochondrial size and position of Golgi complexes varied in different types of spermatogonia. These data show that types of spermatogonia can be differentiated such that these characteristics can be used in functional studies.

Nucleolar variations during the ontogenesis of diploid and tetraploid cyprinid species

Studying the quantitative characteristics of nucleoli in cells of several cyprinid species (Cyprinus carpio, Carassius auratus gibelio, Carassius auratus auratus, Aristichthys nobilis, Ctenopharyngodon idella, Hypophthalmichthys molitrix) has shown a definite programme that controls genetically changes in the nucleolar characteristics during fish ontogenesis. In embryogenesis, the nucleolar activity, which has been estimated from the number and size of nucleoli, decreased; this process was realized dissimilarly in diploid and tetraploid cells. A correlation between the nucleolar characteristics and the body mass of juvenile fish was found. Parameters of the nucleolar activity were closely related to the physiological state of mature organisms and featured tissue specificity, rhythmicity, and high level of heritability.

Effects of chemotherapy on pathologic and biologic characteristics of locally advanced breast cancer.

In 42 patients with locally advanced breast cancer treated with neoadjuvant chemotherapy followed by surgery and radiation therapy, the effects of chemotherapy on tumor architecture, morphometric nuclear and nucleolar characteristics, DNA ploidy, proliferation index measured by mitotic activity index, expression of differentiation antigens, and microvessel density were studied. Pretreatment biopsy specimens were available to compare with mastectomy specimens for 24 patients, and subclavicular biopsy specimens taken before chemotherapy were available for 9 patients. In the remaining patients, fine-needle aspiration was performed before chemotherapy, and morphologic and biologic features of the tumors could be studied only after chemotherapy. In 23 patients, only microscopic tumor or no tumor was left after chemotherapy, and in these patients we observed a characteristic pattern of relatively cellular fibrous tissue with lymphocytic infiltrate, ironloaded macrophages, and, when present, scattered foci of tumor cells in between. We found a reduction in mitotic activity index and in global microvessel density over all the tumors as a group. There was, however, no consistent pattern of changes in nuclear and nucleolar morphometric characteristics, DNA ploidy, and expression of differentiation antigens, and no pathologic or biologic features were predictive for response to chemotherapy.

Prognostic Value of Morphometric Features and the Callender Classification in Uveal Melanomas

Cytomorphometric analysis of uveal melanoma has focused on nucleolar characteristics, although nuclear measurements are easier and more reproducible. In this study, we examine a restricted set of nuclear and nucleolar cytomorphometric variables in the search for the most quantifiable and reproducible prognostic parameter among these and discover a means of assessing the malignancy of tumors within the different Callender subgroups. Uveal melanomas from 94 patients with a minimum follow-up of 5 years underwent nuclear and nucleolar morphometric analysis using a digitized interactive video overlay system. Statistical analysis included univariate and multivariate survival analysis on morphometric variables and classic prognostic parameters (such as Callender cell type, size, etc.). Univariate analysis of cytomorphometric parameters showed the standard deviation of nuclear area (NASD) to be the most significant variable (P < 0.0001: Mantel-Cox, 15.2). This was followed by the difference between nuclear and nucleolar areas (P < 0.002; Mantel-Cox, 9.5), the standard deviation of nucleolar area (SDNA) (P < 0.01; Mantel-Cox, 6.4), and the standard deviation of the shortest nuclear axis (P < 0.02; Mantel-Cox, 6.4). There was a trend for significance with the mean of the ten largest nucleoli (P < 0.07). Multivariate analysis of clinical and cytomorphometric variables showed improvement of prognostic prediction with a combination of the NASD, largest tumor dimension, and glaucoma (P < 0.0001; Mantel-Cox, 26.3). Callender cell type was strongly correlated with the NASD (P < 0.0002), the mean axis ratio of the nuclear area (P < 0.0007), and the mean of the ten largest nucleoli (P < 0.0007). Cytomorphometric analysis of uveal melanoma should include nuclear area and may identify tumors of differing malignancy within the same Callender class.

Differentiating characteristics of myocardial nuclei in cardiomyopathy

Myocardial tissue obtained by endomyocardial biopsy was semiquantitatively evaluated for nuclear and nucleolar characteristics in six groups of patients: patients with normal cardiac function (group 1), doxorubicin-induced cardiomyopathy (group 2), idiopathic cardiomyopathy (group 3), alcoholic cardiomyopathy (group 4), post-viral cardiomyopathy (group 5), and chronic valvular heart disease (group 6). From each patient, ten nuclei containing nucleoli were examined and rated on the basis of the following characteristics: chromatin clumping, number of fibrillar centers per nucleolus, nucleolonemal structure, size of nucleolus, number of nucleoli per nucleus, and stage of nucleolar change. Mean values for the nuclear characteristics in each group were compared with normal values using the unpaired t test. In the doxorubicin treated group there were significantly increased chromatin clumping, decreased fibrillar centers, decreased nucleolonemal structure, and increased numbers of contracted nucleoli (indicating later stage). These changes may be linked to lowered nuclear and nucleolar activity. In the idiopathic and post-viral groups, characteristics were consistent with increased nuclear and nucleolar activity. There were no significant changes from normal in the nucleolar features of either the alcoholic or valvular groups. Further comparisons between groups using one-way analysis of variance and multivariate statistical analysis support the conclusion that there are significant differences in the nuclear and nucleolar characteristics of these groups.

CYTOGENETICS OF A BARLEY × RYE HYBRID

A viable hybrid with a somatic chromosome number of 14 was obtained by crossing Hordeum vulgare L. with Secale cereale L. Chromosome pairing in the hybrid was very low with an average chiasma frequency of 0.22. Phenotypically the hybrid resembled rye, the pollen parent, but the nucleolar characteristics were similar to barley.

Isolation and some characteristics of nucleoli from the brain

A method is described for isolating pure and intact nucleoli from pig and young rat brain cortex using the procedure of Muramatsu and Busch slightly modified. The isolated nucleolar preparation possesses the following properties. (1) Morphological characteristics, as revealed by phase-contrast and electron microscopy, show pure and intact nucleoli. (2) RNA/DNA ratio is about 1.2–1.3. (3) Electrophoretic patterns of nucleolar RNA show evidence of high molecular RNA (45 S, 36 S, 32 S and 28 S), unlike those of extranucleolar RNA (28 S and 18 S). (4) Polyacrylamide gel electrophoresis of 8M urea buffer soluble proteins shows evidence of specific patterns. (5) The nucleolar preparation exhibits RNA synthetic activity. (6) The nucleolar preparation has some ribonuclease activity but no glutamic dehydrogenase activity.

Isolation and Some Characteristics of Nucleoli from the Brain

Abstract A method is described for isolating pure and intact nucleoli from pig and young rat brain cortex using the procedure of Muramatsu and Busch slightly modified. The isolated nucleolar preparation possesses the following properties. (1) Morphological characteristics, as revealed by phase-contrast and electron microscopy, show pure and intact nucleoli. (2) RNA/DNA ratio is about 1.2–1.3. (3) Electrophoretic patterns of nucleolar RNA show evidence of high molecular RNA (45 S, 36 S, 32 S and 28 S), unlike those of extranucleolar RNA (28 S and 18 S). (4) Polyacrylamide gel electrophoresis of 8M urea buffer soluble proteins shows evidence of specific patterns. (5) The nucleolar preparation exhibits RNA synthetic activity. (6) The nucleolar preparation has some ribonuclease activity but no glutamic dehydrogenase activity.