Türkiye has a remarkable geography with its rich inland water ichthyofauna, where many endemic species are found, the genus Cobitis stands out as an important biodiversity element. This genus includes a group of loaches distributed in Europe, Asia and parts of North Africa, usually living in freshwater environments. There are 22 endemic species of Cobitis in Türkiye, distributed in Central, Western and South-Eastern Anatolia. The chromosomal characteristics of C. battalgilae have not been studied, limiting the scope of biological and ecological studies. We employed Giemsa-, C-, and Ag-NOR staining to characterise the karyotype macrostructure of two species of Cobitis from Türkiye, reporting cytogenetic data for the first time in C. battalgilae and reassessing previously known features of C. turcica. The diploid chromosome number in C. battalgilae and C. turcica was 2n = 50 and similar karyotype structures (8 m + 16 sm + 14 st + 12 a; NF = 74) were observed. In both species, there was a secondary constriction on the short arm of autosomal pair 1. Heteromorphic sex chromosomes were not detected. Constitutive heterochromatin (C-bands) was located in the peri/centromeric regions and differed in intensity on some chromosomes. Active Ag-NORs were detected on both homologues and were associated with heterochromatin on the largest metacentric chromosome pair (no. 1) in both species The cytogenetic findings of these two species supported the close relatedness revealed by phylogenetic analysis based on molecular (mitochondrial genes) data.

Cytogenetic insights into Sosippinae (Araneae, Lycosidae) reveal pronounced diploid number reduction in Aglaoctenus and elevated number of rDNA loci in two unrelated species.

Background: CA 125, as a biomarker, holds promise in a wide range of applications, including risk assessment, early detection, diagnosis, prognosis, monitoring, and therapy. Objectives: To find out expression of AgNOR staining in benign, borderline and malignant epithelial tumors of ovary and its correlation with serum CA 125. Materials and Methods: The cross-sectional analytical study, spanning from March 2018 to July 2020 in the Department of Pathology, Sir Salimullah Medical College, Dhaka included 70 diagnosed cases of ovarian epithelial tumors with known CA 125 status. Paraffin blocks and CA 125 reports were gathered, ensuring ethical practices throughout. Results: The mean CA 125 levels in patients with benign was 61.70±30.75 U/mL, in borderline group 65.0±7.07 U/mL and in malignant cases 463.67±249.48 U/mL. CA 125 level was significantly higher in malignant tumors compared to benign and borderline (p<0.001). There was significant positive correlation between CA 125 with mAgNOR (r=0.821; p=0.001) and with pAgNOR (r=0.853; p=0.001). Conclusion: Elevated CA 125 levels were notably significant in malignant tumors compared to benign and borderline cases. A strong, positive correlation existed between CA 125 and both mAgNOR and pAgNOR. J Enam Med Col 2022; 12(3): 137−142

Introduction: Comparative cytogenetics is a vital approach for diagnosing chromosome abnormalities and identifying species-specific patterns. In this study, chromosomal analysis of three Anatolian endemic Cobitis species was performed: Cobitis bilseli, C. fahireae, and C. turcica. Methods: Conventional cytogenetic techniques such as Giemsa staining, C-banding, and Ag-NOR staining were applied, followed by measurements of chromosome arm lengths including analysis of the measured data. Results: The diploid chromosome number, 2n = 50, was determined for all three species. The karyotype formulas were as follows: four pairs of metacentric, 5 pairs of submetacentric, and 16 pairs of subtelo-telocentric chromosomes in C. bilseli; 11 pairs of metacentric, 7 pairs of submetacentric, and 7 pairs of subtelo-telocentric chromosomes in C. fahireae; and 4 pairs of metacentric, 4 pairs of submetacentric, and 17 pairs of subtelo-telocentric chromosomes in C. turcica. Dark C-bands were observed on the pericentromeres of nearly all chromosomes in C. bilseli and C. turcica, whereas light C-bands appeared on the pericentromeres of some chromosomes in C. fahireae. Silver-stained metaphases revealed signals on the short arm of a submetacentric chromosome pair in C. fahireae (each homologous chromosome carries one signal), while in C. bilseli and C. turcica, Ag-NOR signals were detected on the long arm of a single metacentric chromosome (only one homologous chromosome carries the signal, and the signal-carrying chromosome is the largest chromosome in the karyotype). Conclusion: This study provides new cytogenetic data consistent with the phylogenetic distances between the studied species, indicating that pericentric inversions and/or translocations govern the formation of Cobitis karyotypes. Introduction: Comparative cytogenetics is a vital approach for diagnosing chromosome abnormalities and identifying species-specific patterns. In this study, chromosomal analysis of three Anatolian endemic Cobitis species was performed: Cobitis bilseli, C. fahireae, and C. turcica. Methods: Conventional cytogenetic techniques such as Giemsa staining, C-banding, and Ag-NOR staining were applied, followed by measurements of chromosome arm lengths including analysis of the measured data. Results: The diploid chromosome number, 2n = 50, was determined for all three species. The karyotype formulas were as follows: four pairs of metacentric, 5 pairs of submetacentric, and 16 pairs of subtelo-telocentric chromosomes in C. bilseli; 11 pairs of metacentric, 7 pairs of submetacentric, and 7 pairs of subtelo-telocentric chromosomes in C. fahireae; and 4 pairs of metacentric, 4 pairs of submetacentric, and 17 pairs of subtelo-telocentric chromosomes in C. turcica. Dark C-bands were observed on the pericentromeres of nearly all chromosomes in C. bilseli and C. turcica, whereas light C-bands appeared on the pericentromeres of some chromosomes in C. fahireae. Silver-stained metaphases revealed signals on the short arm of a submetacentric chromosome pair in C. fahireae (each homologous chromosome carries one signal), while in C. bilseli and C. turcica, Ag-NOR signals were detected on the long arm of a single metacentric chromosome (only one homologous chromosome carries the signal, and the signal-carrying chromosome is the largest chromosome in the karyotype). Conclusion: This study provides new cytogenetic data consistent with the phylogenetic distances between the studied species, indicating that pericentric inversions and/or translocations govern the formation of Cobitis karyotypes.

Natural product research has an exciting and glorious past that spans over millennia. Accordingly, natural products mediated inhibition of carcinogenesis by mechanistic modulation of deregulated signaling pathways has revolutionized the field of translational oncology. Lycium barbarum has antioxidant and anticarcinogenic effects. The antioxidant activity of the extract and its effect on Ehrlich ascites tumor (EAT) were investigated using in vivo and in vitro techniques. EAT cells were injected into Balb/C mice to create stock mice. EAT cells withdrawn from stock mice were used in equal volumes in the studies. The in vivo study consisted of control and treatment groups (200 mg/kg fractions above and below 50 kDa of extracts). The liver tissues were evaluated for histopathological (H&E), DNA damage (Comet assay), and proliferation (AgNOR staining) status. The in vitro study consisted of control and treatment groups (1500 and 2000 µg/ml of extracts). Cell viability and apoptosis were evaluated. As a result, a decrease in the adhesion of EAT cells, and decreased DNA damage were observed in mice intraperitoneally administered with the fractions of Lycium barbarum. The extracts both below and above 50 kDa increased apoptotic death in cancer cells. The extract above 50 kDa was more active than those below 50 kDa. Lycium barbarum consumption may be effectual in preventing cancer formation and slowing the progression of cancer.

Background/Aim: Cancer has become a prevalent disease, emerging as one of the major chronic health issues today. Currently, common treatments against cancer include chemotherapy, radiotherapy, surgery, and the use of chemically synthesized drugs. However, despite significant advancements in diagnostic methods and treatments, drug resistance and metastasis remain primary hurdles to successful cancer therapy. Consequently, attention has been shifted towards exploring alternative treatments and therapies against cancer. This study sought to examine the time and dose-dependent effects of blueberry (Vaccinium myrtillus L) on MDA-MB-231 cell lines. Methods: The study used the MDA-MB231 breast cancer cell line. We established three groups: control, 40 µl/ml bilberry, and 80 µl/ml bilberry, which were incubated at 37°C and 5% CO2 for 24 and 48 h, respectively. After incubation, we examined the viability, apoptosis, and cell cycle of MDA-MB-231 cells with the Muse Cell Analyzer and assessed the status of nucleolar organizer region (NOR) proteins via silver nitrate (AgNOR) staining. Results: Bilberry extracts were found to enhance apoptosis and exhibit a cytotoxic effect, thereby reducing cell proliferation in MDA-MB-231 cells after 24 and 48 h of culture. There was notably increased apoptosis at concentrations of 40 µl and 80 µl. Moreover, after 48 h of incubation, a significant difference emerged between the control and 40 µg/ml bilberry samples, notably in the average AgNOR count and the total AgNOR area/total nuclear area ratio. Conclusion: Our study suggests that blueberries may be a potential therapeutic candidate for cancer treatment, thereby potentially enriching cancer research.

Impaired rRNA Synthesis Contributes to BCL-2 Induced Chemoresistance in Diffuse Large B-Cell Lymphoma

An exact identification of malignant cells in fluid by cytological examination is a well-known diagnostic challenge. One of the common problems is to distinguish reactive mesothelial cells from malignant cells. Conventional smears reported as ‘suspicious for malignancy’ indicate that the suspicious cases could not be classified with certainty as to whether they were reactive mesothelial cells or malignant cells. It poses problem in clinical staging of tumor, treatment and prognosis of malignancy. The purpose of the study was to determine the role of modified method of AgNOR staining in the evaluation of benign and malignant effusions. This cross-sectional study was conducted in the Department of Pathology, BIRDEM General Hospital, Dhaka, from July 2019 - June 2021. A total of 115 cases of effusion were included. All the samples were centrifuged and then smears were prepared from the deposit followed by staining with Hematoxylin & Eosin stain, Papanicolaou stain and AgNOR stain. At first the diagnosis was made on conventional smear method. Then the findings were compared and analyzed by modified AgNOR staining method. In malignant cells, the mean AgNOR count was 5.59±1.05 (±SD) and the AgNORs were multiple and irregular in shape. On the other hand, in benign cells the AgNORs were comparatively larger, single dots with a mean count of 1.31±0.48.The AgNOR count method has definite role in differentiating benign from malignant effusion. This method has supportive value which can be utilized in differentiating malignant effusions from the benign ones, especially in suspicious cases.

The family Cyprinidae is the largest freshwater fish group with 377 genera and over 3,000 described species. However, this group of fish has very limited cytogenetics and advanced molecular cytogenetics information. Therefore, in this study the karyotypes and other chromosomal characteristics of 15 species in the tribe Systomini (Cyprininae) were examined using Ag-NOR staining along with fluorescence in situ hybridization (5S and 18S rDNA). All species share a similar karyotype (2n = 50; NF = 88-100) in both sexes and no differentiated sex chromosome was observed. Chromosomes bearing NOR sites ranged from one to four pairs among the species, mostly mapped adjacent to telomeres in the short arms of distinct pairs in all analyzed species. This difference indicates an extensive rearrangement of chromosomes including genomic differences. The use of the 5S and 18S rDNA probe confirmed the Ag-NOR sites interstitially located in the telomeric regions of distinct chromosomes, characterizing an interspecies variation of these sites. In most of its analyzed species, the signals of 18S rDNA probe corresponded to the Ag-NOR regions, except in Barbonymus altus, B. gonionotus, B. schwanenfeldii and Puntius brevis having these signals on the same as Ag-NOR regions and other sites.

We present a comparative chromosome study of several taxa of the Malagasy ground geckos of the Paroedura bastardi and P. picta species groups. We employed a preliminary molecular analysis using a trait of the mitochondrial 16S rRNA gene (of about 570 bp) to assess the taxonomic status of the samples studied and a cytogenetic analysis with standard karyotyping (5% Giemsa solution), silver staining (Ag-NOR staining) and sequential C-banding (C-banding + Giemsa and + fluorochromes). Our results show that all the taxa studied of the P. bastardi group (P. ibityensis, P. rennerae and P. cf. guibeae) have a similar karyotype composed of 2n = 34 chromosomes, with two metacentric pairs (1 and 3) and all other pairs being acrocentric. Chromosome diversification in the P. bastardi group was mainly linked to the diversification of heteromorphic sex chromosome systems (ZZ/ZW) in P. ibityensis and P. rennerae, while no heteromorphic sex chromosome pair was found in P. cf. guibeae. The two taxa investigated of the P. picta species group (here named P. picta and P. cf. picta based on molecular data) showed the same chromosome number of 2n = 36, mostly acrocentric elements, but differed in the number of metacentric elements, probably as a result of an inversion at chromosome pair 2. We highlight that the genus Paroedura is characterized by the independent diversification of heterogametic sex chromosomes in different evolutionary lineages and, similarly to other phylogenetically related gecko genera, by a progressive formation of a biarmed element by means of tandem fusions and inversions of distinct pairs.

Giant cell granuloma is a local nonneoplastic lesion that is divided into two categories, based on its site of occurrence: Central and peripheral giant cell granuloma. Central giant cell granuloma is an intraosseous lesion that has a tendency to recure even in surgically treated cases. Several studies have proven that there is an association between different lesions clinical behavior and their histological features. The aim of this study was to evaluate the expression of AgNOR and Ki67 in lesions with and without recurrency. Files and records of 35 patients who had been histologically diagnosed with central giant cell granuloma were investigated. Histological features were studied after performing AgNOR staining and Ki67 marker. The data were analyzed by chi-square, Fisher, and T-test. Acquired data indicated that the count of AgNOR staining and Ki67 marker was significantly higher in lesions with recurrency than the lesions with no recurrency. The same results were attained from Ki67 intensity. The current study indicated that AgNOR staining and Ki67 marker have prognostic value in predicting recurrency of central giant cell granuloma lesions.

The karyological characteristics of nearly half of the Pseudophoxinus species in Türkiye were determined. In this study, it is planned to determine the karyological characteristics of P. anatolicus, which is common in Beyşehir Lake, specimens were caught from the coast at Çiftlik village. The captured specimen were karyological analysed and Giemsa staining, C-banding and Ag-NOR staining were applied to the slides that obtained. The chromosome set of this species consists of 12 pairs of metacentric, eight pairs of submetacentric, two pairs of subtelocentric and three pairs of acrocentric chromosomes. Dark and slightly C-bands were observed in the centromeric regions of some chromosomes. Active Ag-NORs were detected in the telomeric region of the short arm of two pairs of chromosomes. Our results are similar to those of other Pseudophoxinus species except for some differences and it was determined that Anatolian minnow has a conserved karyotype like other Pseudophoxinus species.

Fenugreek or Trigonella foenum-graecum L. is a commercially important yet neglected crop of the family Fabaceae, with potent medicinal applications, and can treat several diseases as well. Conventional breeding studies for higher yields of commercial crops largely depend on chromosomal information of the particular species. Despite a number of cytological research being conducted on T. foenum-graecum, a complete characterization of its chromosomes has not been achieved due to the limitations of traditional karyotype analysis methods. A range of chromosomal markers are advantageous to characterize at full extent and identify individual chromosomes rather than relying on only physical metrics. Thus, in this study, in addition to giemsa staining, other approaches like fluorochrome and silver staining were used for the precise karyomorphological analysis of this species. Enzyme maceration and air drying (EMA) based fluorochrome banding with GC-specific stain Chromomycin A3 (CMA), and AT-specific stain 4’,6-diamidino-2-phenylindole (DAPI) applied for the first time for chromosome characterization. The results showed 2n = 16 chromosomes in metaphase cells, with karyotype formula of 2m+6sm. The unique banding pattern observed in the CMA/DAPI and AgNOR staining highlights the AT and GC-rich regions as well as the nucleolar organizer regions (NORs). All this crucial information can further assist in conducting breeding studies of more precision with simultaneously encouraging similar studies that need to be done in other unexploited species of importance.

This contribution provides the first karyotype description of Hemidactylus mercatorius and discusses the interspecific chromosome diversification in the genus. Chromosomal analysis was performed on samples from different Malagasy populations using standard karyotyping, Ag-NOR staining, and banding methods (sequential C-banding + Giemsa, + Chromomycin A3, +4',6-diamidino-2-phenylindole). Irrespective of sex or sampling locality, H. mercatorius shows a karyotype of 2n = 42 with metacentric (1, 18-21), submetacentric (4), subtelocentric (5, 11), and acrocentric pairs (all the remaining pairs). There was no heteromorphic chromosome pair and no clear distinction between macro- and microchromosomes. NORs were localised close to the centromeres of a medium acrocentric pair (14). Heterochromatic blocks were identified on the telomeric and centromeric regions of most chromosome pairs. A comparison with the karyotype of H. mabouia highlights that the different morphology of several chromosome pairs clearly distinguishes the two species, contrasting the previously proposed synonymy. The differences between the karyotypes of H. mercatorius and H. mabouia concern the number of biarmed and acrocentric elements, suggesting the occurrence of several chromosome inversions. Considering all the available karyotype data on Hemidactylus and its sister genus Cyrtodactylus, it is possible to advance an evolutionary hypothesis on their chromosomal evolution, starting from a common ancestor with 2n = 48 and all acrocentric elements. From this ancestral condition, the karyotype diversification in the two genera has been prevalently characterised by a progressive accumulation of fusions and inversions which have reduced the total chromosome count and increased the number of biarmed chromosomes.

BackgroundMyxofibrosarcoma is a rare malignant soft tissue sarcoma characterised by multiple local recurrence and can become of higher grade with each recurrence. Consequently, myxofibrosarcoma represents a burden for patients, a challenge for clinicians, and an interesting disease to study tumour progression. Currently, few myxofibrosarcoma preclinical models are available.MethodsIn this paper, we present a spontaneously immortalised myxofibrosarcoma patient-derived cell line (MF-R 3). We performed phenotypic characterization through multiple biological assays and analyses: proliferation, clonogenic potential, anchorage-independent growth and colony formation, migration, invasion, AgNOR staining, and ultrastructural evaluation.ResultsMF-R 3 cells match morphologic and phenotypic characteristics of the original tumour as 2D cultures, 3D aggregates, and on the chorioallantoic membrane of chick embryos. Overall results show a clear neoplastic potential of this cell line. Finally, we tested MF-R 3 sensitivity to anthracyclines in 2D and 3D conditions finding a good response to these drugs.ConclusionsIn conclusion, we established a novel patient-derived myxofibrosarcoma cell line that, together with the few others available, could serve as an important model for studying the molecular pathogenesis of myxofibrosarcoma and for testing new drugs and therapeutic strategies in diverse experimental settings.

The monkey goby, Neogobius fluviatilis (Pallas, 1814) that distributed in Türkiye was studied cytogenetically for the first time. In this context, diploid chromosome number, chromosome morphology and also chromosomal banding properties (C-banding and Ag-NOR staining) of N. fluviatilis were revealed out. Chromosome slides were prepared from head kidney cells according to the air-drying technique. Chromosome slides were observed under the microscope and metaphases were photographed. The chromosomes were measured by digital caliper and karyotype was arranged manually. The diploid chromosome number was found as 46. Karyotype was composed with all uniarmed chromosomes. Fundamental arm number was calculated as 46 too. No heteromorphic sex chromosomes were determined in the karyotype. C-bands were detected on the pericentromeres of almost all chromosomes. Otherwise, two Ag-NORs were found in the silver-stained metaphases. This study revealed out chromosomal properties of N. fluviatilis from Türkiye with conventional cytogenetic techniques. This report may improve the cytogenetic data of the genus Neogobius.

In this study, it was aimed to determine the in vitro effect of the extract obtained from mature broccoli on the breast cancer cell line. MDA-MB231 breast cancer cell line was used in the study. MDA-MB-231 cells were exposed to broccoli extract at 37°C and 5% CO2 for varying durations (24 and 48 hours) and doses (125 and 250 µl/ml).At the end of the incubation period, viability, apoptosis, cell cycle and AgNOR protein status of MDA-MB-231 cells were examined in the Muse Cell Analyzer. In the groups containing broccoli extract, a decrease in the percentage of viable cells and a significant increase in the percentage of early and total apoptosis were observed for both doses compared to the control. In the cell cycle test, the number of cells in the S phase increased in all groups.It was observed that the groups containing broccoli extract slowed down the cell cycle in the transition to the S checkpoint. AgNOR staining results also supported cell cycle and apopitosis, and AgNOR number and TAA/NA ratio decreased in the 125 µl/ml broccoli extract group after 24-48 hours and were found to be statistically significant compared to the control group. It was determined that broccoli increased apoptosis on breast cancer cells by various mechanisms and inhibited cell viability/cell growth. The results were similar to the results of AgNOR protein synthesis. The study showed that the regular and correct consumption of broccoli could be effective in preventing cancer formation and slowing its progression.

Sander lucioperca (pikeperch) is a percid fish species of high commercial value and potential for being aquaculture in Turkey. However, karyological studies are deficient for population of Turkey. So, the aim of this study is to carry out diploid chromosome number, karyotype formula, fundamental arm number and chromosomal banding properties (with C-banding and Ag-NOR staining) of S. lucioperca. Specimens of S. lucioperca were captured from Konya, Turkey and alive specimens carried to the laboratory. Chromosome obtaining was provided by using air-drying technique from the head kidney. Chromosome slides were prepared and banding procedures were applied. Result of the analysis, diploid chromosome number was found as 48 and karyotype of the pikeperch consist of 32 biarmed and 16 uniarmed chromosomes. Constitutive heterochromatin regions were observed on the pericentromeres of some of the chromosomes. Ag-NORs were determined on one pair of submetacentric chromosome. This report is the first that determines chromosomal properties of S. lucioperca from Turkey. This study may contribute the cytogenetic information of this species.

Mazamanemorivaga (Cuvier, 1817) is a gray brocket deer that inhabits the Amazon region. An assessment of previous studies revealed inconsistencies in its current taxonomic classification, suggesting the need for an update in its genus classification. A taxonomic repositioning of this species is proposed through the collection of a specimen from its type locality (French Guiana) with subsequent morphological (coloring pattern, body measurements, and craniometry), cytogenetics (G Band, C Band, conventional Giemsa, Ag-NOR staining, and BAC probe mapping), and molecular phylogenetic analysis (mitochondrial genes Cyt B of 920 bp, COI I of 658 bp, D-loop 610 bp), and comparisons with other specimens of the same taxon, as well as other Neotropical deer species. The morphological and cytogenetic differences between this and other Neotropical Cervidae confirm the taxon as a unique and valid species. The phylogenetic analysis evidenced the basal position of the M.nemorivaga specimens within the Blastocerina clade. This shows early diversification and wide divergence from the other species, suggesting that the taxon should be transferred to a different genus. A taxonomic update of the genus name is proposed through the validation of Passalites Gloger, 1841, with Passalitesnemorivagus (Cuvier, 1817) as the type species. Future research should focus on evaluating the potential existence of other species within the genus Passalites, as suggested in the literature.

Germ cell tumors arise from the covering stem cells on the surface of the embryonic ovary. They make up 20-30% of all types of ovarian tumors. About 95% are benign and present as mature cystic teratoma, and only 5% are malignant. Malignant germ cell tumors of the ovary represent 2.6% of all malignant tumors of the ovary, in contrast to epithelial tumors of the ovary (95%). A high incidence of ovarian malignant germ cell tumors is observed in the first two decades of life. Determining the prognosis of germ cell tumors of the ovary is a complex and problematic issue, and according to the existing literature, various methods are used to determine a more accurate classification of cases. One of them can be used AgNOR staining, which is called one of the means of proliferation assessment in the case of different tumors, as well as in the differentiation of dysplasias and benign and malignant processes. According to our research, it is used to evaluate proliferative activity during different histological differentiation of immature teratomas.