Cytological Markers Research Articles

The accuracy of DNA methylation detection in endometrial cancer screening: A systematic review and meta-analysis.

DNA methylation is the hallmark of early endometrial cancer and can be detected through non-invasive methods. The present study systematically reviewed the efficacy of DNA methylation detection for endometrial cancer screening through exfoliative cytology. A systematic literature review was performed through the following databases from inception to October 7, 2024: PubMed, Embase, and the Cochrane Library. Studies on DNA methylation detection for endometrial cancer screening through exfoliative cytology were included. The study was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. The relevant variables included cytologic sample type, methylated genes, and marker performance (i.e., AUC value, sensitivity, specificity, and the corresponding 95% confidence interval [CI]), as well as the bias risk assessment according to the Cochrane Collaboration tool (Cochrane Intervention Systematic Review Guide 5.1.0). A total of 31 genes were selected from 20 studies as methylation markers for endometrial cancer detection in cytologic specimens. A total of 19 methylation markers for endometrial cancer detection with an area under the curve value from 0.80 to 0.97 were selected from 10 studies. Cytology-based DNA methylation markers are feasible and accurate non-invasive methods for the early detection of endometrial cancer screening in high-risk populations.

Equine Asthma Diagnostics: Review of Influencing Factors and Difficulties in Diagnosing Subclinical Disease

This literature review focuses on diagnostics of equine asthma (EA), possible influencing factors on diagnostic techniques and latest developments in diagnosing horses during EA remission or with subclinical disease. Routine EA diagnostics include a clinical examination of the respiratory system with percussion and auscultation including a rebreathing examination, and clinical pathology including white blood cells and arterial blood gas analysis. Subsequent diagnostics include bronchoscopy to evaluate the amount and viscosity of respiratory secretion, bronchoalveolar lavage, and the cytology of tracheal aspirates (TAs) and bronchoalveolar lavage fluid (BALF). The grading of EA severity is built on respiratory effort at rest, which is increased in severe equine asthma. The inflammatory subtype is based on BALF cytology, while TA cytology helps to rule out previous bacterial infections. Different factors have an impact on the airways regarding the structure of the epithelium, cytology, and inflammatory markers possibly influencing the diagnosis of EA. Short-term exercise increases the total cell count and inflammatory mediators identified in the BALF of human patients, asymptomatic horses, and other species. Other factors involve cold or chlorinated air, long-term training effects, and concurrent additional respiratory disease, in particular exercise-induced pulmonary hemorrhage. As BALF cytology may be unremarkable during EA remission and low-grade disease, exercise tests and other factors stressing the bronchial epithelium may help to diagnose these patients.

CNSC-09. OTOSYPHILIS MIMICKING A VESTIBULAR SCHWANNOMA

Abstract Vestibular schwannomas are common nonmalignant primary brain tumors representing up to 80% of the cerebellopontine angle (CPA) tumors. Their diagnosis is made radiographically in most cases with the presence of an enhancing mass along the 8th nerve complex. The United States is experiencing a surge in the number of recorded syphilis cases with an approximate 80% increase since 2018 according to the CDC, after reaching an all-time low in the year 2000. Syphilis can involve the Central Nervous System (CNS) in its early, late, and latent forms, and in its early stages it typically presents in the form of meningitis which can be accompanied by cranial neuropathies especially of the nerves II, VII, and VIII. We present a case of a 35-year-old man who presented with subacute onset unilateral left-sided hearing loss and dizziness that did not respond to prescribed steroids. Magnetic resonance imaging (MRI) revealed an enhancing mass in the left cerebellopontine angle presumed to be a vestibular schwannoma. The patient was subsequently planned for radiosurgery, however, MRI done during the procedure revealed that the mass was of a smaller size. The patient was referred to neuro-oncology and work up included cerebrospinal fluid (CSF) analysis that showed 3 white blood cells, normal protein and glucose, and negative VDRL. Cytology, flow cytometry, and inflammatory markers were negative. However, the Treponema pallidum antibodies were positive confirming otoshyphilis. The patient was treated accordingly with a course of Penicillin G with near resolution of the enhancement of the 8th nerve complex. His hearing remained impaired on his initial follow up, however, his facial weakness near resolved. This case brings up an important diagnostic question in cases of presumed schwannomas where patients would be otherwise treated solely based on the MRI appearance, especially given the re-emergence of syphilis.

Phylogenetic Relationship Among Cultivated and Wild Species of Genus Macrotyloma Based on Cytology and Molecular Markers

Phylogenetic Relationship Among Cultivated and Wild Species of Genus Macrotyloma Based on Cytology and Molecular Markers

Genetic Diversity Research of Yam Germplasm Resources

Yam (Dioscorea spp.), a versatile economic crop used both as food and medicine, is widely distributed globally, particularly in tropical and subtropical regions. Its abundant germplasm resources provide a crucial foundation for genetic diversity research. This paper reviews the latest advances in the genetic diversity study of yam germplasm resources, focusing on research methods and outcomes based on morphological traits, cytological markers, and molecular markers. By analyzing the application and effectiveness of these research methods, this paper summarizes key findings in yam genetic diversity research and proposes future research directions, aiming to provide theoretical support and technical guidance for yam genetic improvement and germplasm resource conservation.

FSH Therapy in Male Factor Infertility: Evidence and Factors Which Might Predict the Response.

Follicle-stimulating hormone (FSH) administration is applied in the management of subjects affected by hypogonadotropic hypogonadism. Whilst this application is widely recognized and established alone or in combination with human chorionic gonadotropin (hCG), a similar strategy is empirically advocated in idiopathic male factor infertility (MFI). In this setting, FSH therapy has been used to increase sperm quantity, quality, and pregnancy rate when FSH plasma concentrations are below 8 IU/L and when the seminal tract is not obstructed. In the literature, several studies suggested that giving FSH to patients with idiopathic MFI increases sperm count and motility, raising the overall pregnancy rate. However, this efficacy seems to be limited, and about 10-18 men should be treated to achieve one pregnancy. Thus, several papers suggest the need to move from a replacement approach to an overstimulating approach in the management of FSH therapy in idiopathic MFI. To this aim, it is imperative to determine some pharmacologic markers of FSH efficacy. Furthermore, it should be useful in clinical practice to distinguish, before starting the treatment, among patients who might respond or not to FSH treatment. Indeed, previous studies suggest that infertile men who have normal levels of gonadotropins in plasma might not respond to FSH treatment and about 50% of patients might be defined as "non-responders". For these reasons, identifying predictive markers of FSH action in spermatogenesis and clinical markers of response to FSH treatment is a fascinating area of study that might lead to new developments with the aim of achieving personalization of the treatment of male infertility. From this perspective, seminal parameters (i.e., spermatid count), testicular cytology, genetic assessment, and miRNA or protein markers in the future might be used to create a tailored FSH therapy plan. The personalization of FSH treatment is mandatory to minimize side effects, to avoid lost time with ineffective treatments, and to improve the efficacy, predicting the most efficient dose and the duration of the treatment. This narrative review's objective is to discuss the role of the different putative factors which have been proposed to predict the response to FSH treatment in idiopathic infertile men.

The chromatin-associated 53BP1 ortholog, HSR-9, regulates recombinational repair and X chromosome segregation in the Caenorhabditis elegans germ line.

53BP1 plays a crucial role in regulating DNA damage repair pathway choice and checkpoint signaling in somatic cells; however, its role in meiosis has remained enigmatic. In this study, we demonstrate that the Caenorhabditis elegans ortholog of 53BP1, HSR-9, associates with chromatin in both proliferating and meiotic germ cells. Notably, HSR-9 is enriched on the X chromosome pair in pachytene oogenic germ cells. HSR-9 is also present at kinetochores during both mitotic and meiotic divisions but does not appear to be essential for monitoring microtubule-kinetochore attachments or tension. Using cytological markers of different steps in recombinational repair, we found that HSR-9 influences the processing of a subset of meiotic double-stranded breaks into COSA-1-marked crossovers. Additionally, HSR-9 plays a role in meiotic X chromosome segregation under conditions where X chromosomes fail to pair, synapse, and recombine. Together, these results highlight that chromatin-associated HSR-9 has both conserved and unique functions in the regulation of meiotic chromosome behavior.

Different Frequency and Clinical Role for MTAP Loss in Pleural and Peritoneal Mesothelioma.

The objective of this study was to analyze the expression and prognostic role of methylthioadenosine phosphorylase (MTAP) in mesothelioma. MTAP protein expression by immunohistochemistry was analyzed in 113 mesotheliomas (60 pleural and 53 peritoneal), consisting of 36 effusions and 77 surgical specimens. MTAP expression was fully lost in 38 tumors and partially lost in 8 tumors. Loss of expression was significantly more common in effusions compared with biopsies/surgical resection specimens (20/36 vs. 26/77; P =0.017), and in pleural compared with peritoneal mesotheliomas (35/60 vs. 11/53; P <0.001). MTAP performed less robustly than BAP1 in comparative analysis of 57 tumors previously analyzed for expression of the latter protein (46 vs. 25 cases with loss of expression). In survival analysis for 69 patients with partial clinical data, male gender was significantly associated with shorter overall survival (OS; P =0.042), whereas loss of MTAP was associated with a trend for shorter OS ( P =0.058), with no prognostic role for patient age ( P =0.379) or anatomic site ( P =0.381). The association between loss of MTAP and poor OS became significant when survival analysis was limited to patients with pleural mesothelioma ( P =0.018). In conclusion, loss of MTAP expression is more frequent in pleural compared with peritoneal mesothelioma and has limited diagnostic relevance at the latter anatomic site. More frequent loss in effusion specimens suggests a role for this marker in effusion cytology. MTAP loss in pleural mesothelioma is associated with poor survival.

TFE3 -Rearranged PEComa/PEComa-like Neoplasms : Report of 25 New Cases Expanding the Clinicopathologic Spectrum and Highlighting its Association With Prior Exposure to Chemotherapy.

Since their original description as a distinctive neoplastic entity, ~50 TFE3 -rearranged perivascular epithelioid cell tumors (PEComas) have been reported. We herein report 25 new TFE3 -rearranged PEComas and review the published literature to further investigate their clinicopathologic spectrum. Notably, 5 of the 25 cases were associated with a prior history of chemotherapy treatment for cancer. This is in keeping with prior reports, based mainly on small case series, with overall 11% of TFE3 -rearranged PEComas being diagnosed postchemotherapy. The median age of our cohort was 38 years. Most neoplasms demonstrated characteristic features such as nested architecture, epithelioid cytology, HMB45 positive, and muscle marker negative immunophenotype. SFPQ was the most common TFE3 fusion partner present in half of the cases, followed by ASPSCR1 and NONO genes. Four of 7 cases in our cohort with meaningful follow-up presented with or developed systemic metastasis, while over half of the reported cases either recurred locally, metastasized, or caused patient death. Follow-up for the remaining cases was limited (median 18.5 months), suggesting that the prognosis may be worse. Size, mitotic activity, and necrosis were correlated with aggressive behavior. There is little evidence that treatment with MTOR inhibitors, which are beneficial against TSC -mutated PEComas, is effective against TFE3 -rearranged PEComas: only one of 6 reported cases demonstrated disease stabilization. As co-expression of melanocytic and muscle markers, a hallmark of conventional TSC -mutated PEComa is uncommon in the spectrum of TFE3 -rearranged PEComa, an alternative terminology may be more appropriate, such as " TFE3 -rearranged PEComa-like neoplasms," highlighting their distinctive morphologic features and therapeutic implications.

Diagnostic Accuracy of Cancer in Pleural Effusion: A Combination Serum Tumor Markers and Cytology

Abstract Background: Cytological examination of body effusion has an essential role in different tumor staging and research of malignancy. Objectives: It is to study the diagnostic effect of serum tumor markers in differentiation between benign and malignant pleural effusion. Materials and Methods: A retrospective research was done on 39 patients (28 male and 11 female with an age range of 50–77 years old). All samples undergo diagnostic thoracocentesis. The cutoff values of serum tumor markers (CEA &lt; 5 ng/mL; CA125 &lt;35 U/mL; CA19-9 &lt; 37 U/mL; and CA15-3 &lt; 31 U/mL). Results: Thirty-nine cases, resulting in 26 (67%) benign pleural effusions and 13 (33%) malignant pleural effusions, mean ± SD of age was higher malignant pleural effusion than benign type. The sensitivity of cytology was 33%. Malignant pleural effusion tended to have higher serum marker tumor concentrations. A comparison association between benign and malignant pleural effusion revealed statistically significant only in CEA. Most areas under the curve had moderate values that ranged between 0.6 for CA125, CA15.3, and other combined tumor markers, except for CEA, which had a good AUC (0.799). The best efficacy of a single tumor marker is CEA with a cutoff value of 2.05 ng/mL; at this cutoff value, the sensitivity, specificity, and accuracy were 69.2%, 88.5%, and 82.1%, respectively. Conclusion: Single serum tumor markers showed moderate sensitivity and low specificity; their combination does not result in to increase level of sensitivity and specificity to a good and dependent level for diagnosis of the malignant pleural effusion. Cytology and serum tumor markers have aided role in diagnosis. Thoracoscopy and pleural biopsy remain dependent diagnostic tools for malignant pleural effusion.

Evaluating the comprehensive diagnosis efficiency of lung cancer, including measurement of SHOX2 and RASSF1A gene methylation

Methylation of the promoters of SHOX2 and RASSF1A (LungMe®) exhibits promise as a potential molecular biomarker for diagnosing lung cancer. This study sought to assess the aberrant methylation of SHOX2 and RASSF1A in broncho-exfoliated cells (BEC) and compare it with conventional cytology, histology examination, immunohistochemistry, and serum tumor markers to evaluate the overall diagnostic efficiency for lung cancer. This study recruited 240 patients, including 185 malignant cases and 55 benign cases. In our observation, we noted a slight reduction in the detection sensitivity, however, the ΔCt method exhibited a significant enhancement in specificity when compared to Ct judgment. Consequently, the ΔCt method proves to be a more appropriate approach for interpreting methylation results. The diagnostic sensitivity of cytology and histology was in ranged from 20.0%-35.1% and 42.9%-80%, respectively, while the positive detection rate of LungMe® methylation ranged from 70.0% to 100%. Additionally, our findings indicate a higher prevalence of SHOX2( +) among patients exhibiting medium and high expression of Ki67 (P < 0.01), as opposed to those with low expression of Ki67, but RASSF1A methylation did not show this phenomenon (P = 0.35). Furthermore, CEA, SCCA, and CYFRA21-1 showed positive detection rates of 48.8%, 26.2%, and 55.8%, respectively. Finally, we present a comprehensive lung cancer diagnostic work-up, including LumgMe® methylation. The combined analysis of SHOX2 and RASSF1A methylation serves as a powerful complement and extension to conventional methods, enhancing the accuracy of a lung cancer diagnosis with satisfactory sensitivity and specificity.

Development and application of whole-chromosome painting of chromosomes 7A and 8A of Arachis duranensis based on chromosome-specific single-copy oligonucleotides.

For peanut, the lack of stable cytological markers is a barrier to tracking specific chromosomes, elucidating the genetic relationships between genomes and identifying chromosomal variations. Chromosome mapping using single-copy oligonucleotide (oligo) probe libraries has unique advantages for identifying homologous chromosomes and chromosomal rearrangements. In this study, we developed two whole-chromosome single-copy oligo probe libraries, LS-7A and LS-8A, based on the reference genome sequences of chromosomes 7A and 8A of Arachis duranensis. Fluorescence in situ hybridization (FISH) analysis confirmed that the libraries could specifically paint chromosomes 7 and 8. In addition, sequential FISH and electronic localization of LS-7A and LS-8A in A. duranensis (AA) and A. ipaensis (BB) showed that chromosomes 7A and 8A contained translocations and inversions relative to chromosomes 7B and 8B. Analysis of the chromosomes of wild Arachis species using LS-8A confirmed that this library could accurately and effectively identify A genome species. Finally, LS-7A and LS-8A were used to paint the chromosomes of interspecific hybrids and their progenies, which verified the authenticity of the interspecific hybrids and identified a disomic addition line. This study provides a model for developing specific oligo probes to identify the structural variations of other chromosomes in Arachis and demonstrates the practical utility of LS-7A and LS-8A.

Ac-DEVD-CHO (caspase-3/DEVDase inhibitor) suppresses self-incompatibility–induced programmed cell death in the pollen tubes of petunia (Petunia hybrida E. Vilm.)

Programmed cell death (PCD) is relevant to many aspects in the growth and development of a plant organism. In their reproduction, many flowering plant species possess self-incompatibility (SI), that is an intraspecific reproductive barrier, which is a genetic mechanism ensuring the avoidance of inbreeding depression by preventing self-pollination. This phenomenon enhances intraspecific variation; however, SI is rather a hindrance for some fruit plant species (such as plum, cherry, and peer trees) rather than an advantage in farming. PCD is a factor of the S-RNase–based SI in Petunia hybrida E. Vilm. The growth of self-incompatible pollen tubes (PTs) is arrested with an increase in the activity of caspase-like proteases during the first hours after pollination so that all traits of PCD—plasma membrane integrity damage, DNA degradation/disintegration, and damage of PT structural organization (absence of vacuoles, turgor disturbance, and separation of cell plasma membrane from the cell wall)—are observable by the moment of PT growth arrest. We succeeded in discovering an additional cytological PCD marker, namely, the formation of ricinosomes in self-incompatible PTs at early stages of PCD. SI is removable by treating petunia stigmas with Acetyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO), an inhibitor of caspase-3/DEVDase, 2 h before a self-incompatible pollination. In this process, the level of caspase-3-like protease activity was low, DNA degradation was absent, PTs grew to the ovary, fertilization was successful, and full-fledged seeds were formed.

Astrocyte-Neuron Interactions in Substance Use Disorders.

Engagement of astrocytes within the brain's reward circuitry has been apparent for approximately 30years, when noncontingent drug administration was observed to lead to cytological markers of reactive astrocytes. Since that time, advanced approaches in rodent behavior and astrocyte monitoring have revealed complex interactions between astrocytes with drug type, animal sex, brain region, and dose and duration of drug administration. A number of studies now collectively reveal that rodent drug self-administration followed by prolonged abstinence results in decreased features of structure and synaptic colocalization of astrocytes. In addition, stimulation of astrocytes in the nucleus accumbens with DREADD receptors or pharmacological compounds opposes drug-seeking behavior. These findings provide a clear path for ongoing investigation into astrocytes as mediators of drug action in the brain and underscore the potential therapeutic utility of astrocytes in the regulation of drug craving and relapse vulnerability.

Bleomycin-treated myoblasts undergo p21-associated cellular senescence and have severely impaired differentiation.

As we age, the ability to regenerate and repair skeletal muscle damage declines, partially due to increasing dysfunction of muscle resident stem cells-satellite cells (SC). Recent evidence implicates cellular senescence, which is the irreversible arrest of proliferation, as a potentiator of SC impairment during aging. However, little is known about the role of senescence in SC, and there is a large discrepancy in senescence classification within skeletal muscle. The purpose of this study was to develop a model of senescence in skeletal muscle myoblasts and identify how common senescence-associated biomarkers respond. Low-passage C2C12 myoblasts were treated with bleomycin or vehicle and then evaluated for cytological and molecular senescence markers, proliferation status, cell cycle kinetics, and differentiation potential. Bleomycin treatment caused double-stranded DNA breaks, which upregulated p21 mRNA and protein, potentially through NF-κB and senescence-associated super enhancer (SASE) signaling (p < 0.01). Consequently, cell proliferation was abruptly halted due to G2/M-phase arrest (p < 0.01). Bleomycin-treated myoblasts displayed greater senescence-associated β-galactosidase staining (p < 0.01), which increased over several days. These myoblasts remained senescent following 6days of differentiation and had significant impairments in myotube formation (p < 0.01). Furthermore, our results show that senescence can be maintained despite the lack of p16 gene expression in C2C12 myoblasts. In conclusion, bleomycin treatment provides a valid model of damage-induced senescence that was associated with elevated p21, reduced myoblast proliferation, and aberrant cell cycle kinetics, while confirming that a multi-marker approach is needed for the accurate classification of senescence within skeletal muscle.

Retinol palmitate in management of chronic Steven-Johnson Syndrome with ocular surface keratinization

PurposeTo study the outcomes of topical Retinol Palmitate ophthalmic solution in chronic Stevens-Johnson Syndrome with ocular surface keratinisation. MethodsIt was a comparative interventional study conducted at Rajendra Prasad Centre for Ophthalmic Sciences, Delhi, India from 2020 to 2022 evaluating outcomes of addition of topical Retinol Palmitate to conventional treatment objectively as well as subjectively from baseline up to 12 weeks. ResultsA statistically significant improvement was seen in patients who received topical Retinol palmitate at 12 weeks in terms of Schirmer-1 test(p=<0.01), tear prism height on ASOCT(p = 0.02), Rose Bengal staining score of cornea(p = 0.01) and conjunctiva (p < 0.01), reduction of ocular surface keratinisation on impression cytology(p = 0.01) and subjective evaluation using OSDI questionnaire(p = 0.04).Impression cytology revealed goblet cells in Retinol palmitate group at 1 week after initiation of therapy, which increased further at 1 month follow up but reduced at 3 months. No goblet cells were seen in control group at any follow-up. No significant difference was noted between the two groups in terms of visual acuity, tear film breakup time, inflammatory cells on impression cytology and inflammatory markers in tears. ConclusionTopical Retinol palmitate is a safe and effective drug in cases of chronic SJS with ocular surface keratinisation. It has the potential to reverse keratinisation of the ocular surface and promote development of goblet cells. However, the survival of goblet cells is not long lasting.

Morphological to Molecular Markers: Plant Genetic Diversity Studies in Walnut (Juglans regia L.)—A Review

Persian walnut populations have tremendous morphological and allelic diversity in their germplasm due to heavy outcrossing and years of seed multiplication. These variations are assessed by morphological, cytological, biochemical, and molecular markers. Various researchers have used different tree, foliage, flower, nut, and kernel traits to evaluate morphological/phenotypic diversity. In walnut, morphological indices are first taken into consideration to describe and classify the germplasm, but the environment influences them. In comparison, DNA-based markers can detect genetic diversity at any stage of plant development and have been shown to be a potential tool for assessing variation at the DNA level and deciphering genetic relationships within and between species. Microsatellites are very powerful and informative among DNA-based markers in studying genetic relationships and genetic identity at different levels. They are neutral, highly frequent, uniformly distributed, hypervariable, codominant, highly reproducible, produce many alleles per locus, and require a small amount of DNA for analysis. Current breeding objectives can be achieved by selecting superior genotypes from the germplasm, supplemented by molecular characterization in the selection of parents for each breeding program. Therefore, the use of morphological and molecular markers is recommended for efficient exploration and utilization of germplasm resources and to improve diversity among genetic resources. The published literature on morphological and molecular markers, especially simple sequence repeats (SSRs), is presented in this review to provide current insights into the level of genetic diversity in walnut.

Hemocytes of Yesso scallop characterized by cytological, molecular marker, and functional analyses

Bivalve hemocytes have pivotal role as cellular biodefense. However, no information is available for cytological parameters, marker gene and function of the hemocytes in Yesso scallop, a commercially important aquaculture species worldwide. Due to their extremely strong cell aggregation ability, the scallop hemocytes were not able to assess as a single cell so far. In the present study, we established methodologies for studying the hemocytes of Yesso scallop, assessed cell morphology, measured seasonal fluctuation, and analyzed transcriptomes and cellular behavior during the immune response. Our results showed that the Yesso scallop possesses a single type of leukocyte-type hemocytes similar to other bivalve granulocytes circulating at an average of 1 × 107 cells/ml throughout the year. In addition, we identified five molecular marker genes specific to the scallop hemocytes. These hemocyte markers enabled us to precisely detect the hemocyte localization. Using these markers, we confirmed that tissue transplantation can experimentally induce an immune response, leading to the mobilization of circulating hemocytes for encapsulation. This study provides a comprehensive understanding of scallop hemocytes and their role in the cellular biodefense system of bivalves and various methods for cytological analysis.

Phospho PTEN mediated dephosphorylation of mitotic kinase PLK1 and Aurora Kinase A prevents aneuploidy and preserves genomic stability

PTEN, dual phosphatase tumor suppressor protein, is found to be frequently mutated in various cancers. Post-translational modification of PTEN is important for its sub-cellular localization and catalytic functions. But how these modifications affect cytological damage and aneuploidy is not studied in detail. We focus on the role of phosphatase activity along with C-terminal phosphorylation of PTEN in perspective of cytological damage like micronucleus, nuclear bud, and nuclear bridge formation. Our data suggest that wild-type PTEN, but not phospho-mutant PTEN significantly reduces cytological damage in PTEN null PC3 cells. In case of phosphatase-dead PTEN, cytological damage markers are increased during 24h recovery after DNA damage. When we use phosphorylation and phosphatase-dead dual mutant PTEN, the extent of different cytological DNA damage parameters are similar to phosphatase-dead PTEN. We also find that both of those activities are essential for maintaining chromosome numbers. PTEN null cells exhibit significantly aberrant γ-tubulin pole formation during metaphase. Interestingly, we observed that p-PTEN localized to spindle poles along with PLK1 and Aurora Kinase A. Further depletion of phosphorylation and phosphatase activity of PTEN increases the expression of p-Aurora Kinase A (T288) and p-PLK1 (T210), compared to cells expressing wild-type PTEN. Again, wild-type PTEN but not phosphorylation-dead mutant is able to physically interact with PLK1 and Aurora Kinase A. Thus, our study suggests that the phosphorylation-dependent interaction of PTEN with PLK1 and Aurora Kinase A causes dephosphorylation of those mitotic kinases and by lowering their hyperphosphorylation status, PTEN prevents aberrant chromosome segregation in metaphase.

Tandem Repeat DNA Provides Many Cytological Markers for Hybrid Zone Analysis in Two Subspecies of the Grasshopper Chorthippus parallelus.

Recent advances in next generation sequencing (NGS) have greatly increased our understanding of non-coding tandem repeat (TR) DNA. Here we show how TR DNA can be useful for the study of hybrid zones (HZ), as it serves as a marker to identify introgression in areas where two biological entities come in contact. We used Illumina libraries to analyse two subspecies of the grasshopper Chorthippus parallelus, which currently form a HZ in the Pyrenees. We retrieved a total of 152 TR sequences, and used fluorescent in situ hybridization (FISH) to map 77 families in purebred individuals from both subspecies. Our analysis revealed 50 TR families that could serve as markers for analysis of this HZ, using FISH. Differential TR bands were unevenly distributed between chromosomes and subspecies. Some of these TR families yielded FISH bands in only one of the subspecies, suggesting the amplification of these TR families after the geographic separation of the subspecies in the Pleistocene. Our cytological analysis of two TR markers along a transect of the Pyrenean hybrid zone showed asymmetrical introgression of one subspecies into the other, consistent with previous findings using other markers. These results demonstrate the reliability of TR-band markers for hybrid zone studies.