Cytomorphological Changes Research Articles

Biodegradable rice bran insoluble dietary fiber-chitosan blend films: Characterization and potential applications

A composite film of insoluble dietary fiber (IDF) from rice bran (RB) blended with chitosan (IDF-CS film) was prepared and characterized. The thermal stability, thickness, tensile strength, colour, water vapour permeability (WVP) and oil permeability (PO) of the film were analysed. SEM micrographs revealed the porous morphology of the film. XRD spectra suggested an amorphous crystalline nature of the film. FTIR spectra showed a 1600 cm−1 peak in chitosan and dietary fiber, indicating C=O stretching, and TGA showed good thermal stability under various temperatures. The thickness increased to 0.104 mm in the film containing IDF -Chitosan2:2. The tensile strength (TS) was enhanced to 18.420 MPa in the film containing IDF -Chitosan2:2. The elongation at break (EB) was higher (12.65%) in films containing low concentrations of IDF-Chitosan0.5:0.5. WVP was enhanced to 5.856 ×10-12 g·cm/cm2·Pa·s in the film containing IDF 2:2. The prepared film showed excellent soil-degrading properties, with a degradation rate of 96% after 15 days. The IDF-CS film demonstrated a substantial inhibitory potential against Gram-positive bacteria. The scavenging capacities of the degradable film were 96.5% for DPPH and 98.8% for ABTS. Moreover, the viability of cervical HeLa cell line was 10% at 200 μg/ml compared with that of the positive control (4%). Significant cytomorphological changes leading to necrosis in treated cells were observed using light and fluorescence microscopy. The film (IDF-CS) developed is environmentally friendly and could be used as a novel therapeutic material for treating bacterial pathogens, scavenging free radicals, and inhibiting cancer cell proliferation.

Cervical intraepithelial neoplasia and cervical cytology in pregnancy.

Cervical cancer screening during pregnancy presents unique challenges for cytologic interpretation. This review focuses on pregnancy-associated cytomorphological changes and their impact on diagnosis of cervical intraepithelial neoplasia (CIN) and cervical cancer. Pregnancy-induced alterations include navicular cells, hyperplastic endocervical cells, immature metaplastic cells, and occasional decidual cells or trophoblasts. These changes can mimic abnormalities such as koilocytosis, adenocarcinoma in situ, and high-grade squamous intraepithelial lesions, potentially leading to misdiagnosis. Careful attention to nuclear features and awareness of pregnancy-related changes are crucial for correct interpretation. The natural history of CIN during pregnancy shows higher regression rates, particularly for CIN 2, with minimal risk of progression. Management of abnormal cytology follows modified risk-based guidelines to avoid invasive procedures, with treatment typically deferred until postpartum. The findings reported in this review emphasize the importance of considering pregnancy status in cytological interpretation, highlight potential problems, and provide guidance on differentiating benign pregnancy-related changes from true abnormalities. Understanding these nuances is essential for accurate diagnosis and proper management of cervical abnormalities in pregnant women.

HPV-Independent Cervical Adenocarcinoma Discovered by Cytology-Based Screening: A case report of gastric type adenocarcinoma of the uterine cervix from cytopathology to histopathology

Abstract Introduction/Objective In recent years, the relative prevalence of adenocarcinoma has increased, comprising 20–25% of all cervical carcinomas in developed countries. Approximately 15% of cervical adenocarcinomas are unrelated to human papilloma virus (HPV) infection. With the shift toward HPV primary testing, an increased understanding of HPV-independent cervical adenocarcinomas becomes more important. Methods/Case Report This case report demonstrates a rare case of gastric-type adenocarcinoma (GAS) of the cervix discovered during a routine well-woman exam. Patient-reported episodes of vaginal spotting were noted prior to screening. HPV testing by PCR on the initial pap smear was negative. Cytology findings raised concerns about endocervical glandular cell abnormalities. Subsequent cervical biopsy revealed GAS, a finding supported by positive CK7, MUC5AC, CDX2, and aberrant (mutant) p53 expression by immunohistochemistry. HPV independence was further confirmed by negative results on mRNA ISH for high-risk HPV and HPV16/18. Results (if a Case Study enter NA) NA Conclusion Here we discuss the impact of cytology-based screening on HPV-independent cervical cancers, with focus on adenocarcinoma. While the move to primary HPV screening has been shown to be effective at identifying HPV-related neoplasms, it ineffectively addresses HPV-independent malignancies. In this particular population, cytology-based screening remains indispensable. The described case demonstrated subtle cytologic features consistent with endocervical glandular abnormality, ultimately correlating with subsequent histologic findings of HPV-independent GAS. Negative HPV results combined with subtle cytomorphologic changes displayed by some adenocarcinomas on PAP smear could present a significant diagnostic pitfall. Additionally, p16+ expression attributable to aberrant p53 pathways (rather than HPV) may further contribute to potential diagnostic errors. While further research into best practices for early detection of HPV-independent cervical neoplasias should be conducted, this team suggests both clinical survey and referral for cytologic screening following reports of abnormal vaginal bleeding and/or watery discharge and screener education focused on cervical GAS.

Dynamics of indicators of the functional state of the body and cytomorphological changes in wrestlers during the performance at the championship of Ukraine in belt wrestling «Alysh»

Dynamics of indicators of the functional state of the body and cytomorphological changes in wrestlers during the performance at the championship of Ukraine in belt wrestling «Alysh»

Green synthesis of ultrafine gold nanoparticles from sweet flag (Acorus calamus) for effective anti-leukemic, anti-urolithiatic and in silico docking studies

Gold nanoparticles (AuNPs) are emerging effective biomaterials which play a vital role in the biodiversity research area due to their biocompatibility, and less toxic nature. The AuNPs of various sizes and shapes have vibrant optical features that can be utilized for a variety of imaging and therapeutic applications. Acorus calamus (A. calamus) has been used in traditional medical systems as a general tonic for centuries. It is thought to have potential benefits in the treatment of digestive disorders and respiratory problems. In Kerala, newborns were given a nanogram of (A. calamus) rhizome paste to increase oral cleanliness and improve their health and immunity. This prompted us to produce AuNPs by A. calamus and to explore their concealed pharmacological properties. In this work, we used a simple sonication method to synthesise blue-coloured AuNPs using sweet flag (A. calamus) extract, which acted as a stabilising and reducing agent at ambient temperature. The formation and morphology of the AC-AuNPs was confirmed by XRD, TEM, EDX, FT-IR and UV–vis spectral techniques. Anti-leukemic and anti-urolithiatic activities of the AC-AuNPs were also investigated. The anti-leukemic results revealed cytomorphological changes like cancer cell membrane lyses and coiling in Human myelogenous leukemia cells (K562) and exhibited IC50 at a concentration of 41.45 µg/mL. Furthermore, in vitro DNA binding assay of AC-AuNPs with calf-thymus were conducted. The anti-urolithiatic activity of AC-AuNPs suggests a dose-dependent suppression of calcium oxalate method (COM) crystals nucleation and aggregation. These results suggest that AC-AuNPs have potent antileukemic and anti-urolithiatic activities. In silico docking results revealed that eugenol, α-asarone and calamenene possess good binding scores with myeloid cell surface antigen (CD33) and estrogen receptors, respectively. In vitro anticancer and DNA binding studies have endorsed the capping nature of AC-AuNPs by these phytoconstituents, which is validated by docking analysis.

Cytomorphological Changes in Patients Undergoing Taxanes Formation in Breast Cancer at Oncology and Cancer Research Center in Shendi Town, Sudan

Background: Breast cancer is the most commonly diagnosed cancer in women worldwide and the most frequent cause of cancer death in women in low-income countries. Cytotoxic chemotherapy drugs called taxanes alter the cellular mechanisms that allow cancer cells to divide their microtubules. In a typical cell cycle, microtubules are formed by the cell at the start of division and disassembled after the cell stops dividing. Taxanes stabilize the microtubules, preventing them from breaking down normally. This causes the cancer cells to stop dividing, potentially slowing the growth of cancer or killing the cells. Taxane causes neutropenia and thrombocytopenia, as well as fatigue, nausea and vomiting, hair loss, diarrhea, mouth ulcers, and muscle pain. According to the literature, chemotherapy causes nuclear atypia and other cellular changes in buccal mucosa cells. There was no published data regarding the effect of taxane on buccal mucosa cells. This study aimed to detect cytomorphological changes among breast cancer patients receiving taxane chemotherapy. Materials and methods: This is a comparative cross-sectional study conducted during the period from December 2022 to February 2024, involved 50 buccal smears from breast cancer patients included as a case group versus 50 buccal smears from apparently healthy females. All smears were processed through conventional cytological methods. Results: Forty percent of patients have had a family history of breast cancer. We observed that in 80% of patients with advanced age (46 years old and older), our results indicated that there was a significant statistical difference between cases and controls regarding cellular changes in buccal mucosa. We also observed an increase in the significant correlation of cellular changes and the number of taxane doses, as the p value was less than 0.05. Conclusion: We concluded that taxane chemotherapy may cause buccal cellular changes as the incidence of nuclear atypia, infection, and degenerative changes is higher in cases than in controls.

Cytopathological study of pleural effusion in SARS-CoV2 RTPCR positive subjects during pandemic at tertiary care hospital

Introduction and Aim: Respiratory system is the most common system affected by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Review of literature reveals that pleural effusion were seen in one third of cases with Middle East Respiratory Syndrome(MERS) but were rarely reported in SARS –CoV2.This study aims to study the cytomorphological changes in Pleural effusion seen in SARS CoV-2 positive cases, to correlate the cytological findings with radiological CT COVID-19 Reporting and Data System(CORADS)scorings and to categorize pleural effusion as isolated cause of SARS CoV-2 or aggravation of underlying other etiologies. Materials and Methods: Pleural fluid samples of SARS CoV-2 RT PCR positive cases, obtained from patients of all age groups were included in the study. The samples were predominantly from Covid19 wave 1 (March 2020- February 2021) and few from Covid wave 2(March 2021-April 2021). Biochemical analysis, hematological parameters, cytomorphology and associated comorbidity was recorded, and analysis done. Results: Male predominance was observed. Most common cytological and biochemical finding in pleural fluid analysis was Exudative pleural effusion. Predominant cytological features showed lymphocytic effusion. Radiology showed maximum left sided pleural effusion (42%). CORADS score was 6 in majority of cases and effusion as an isolated cause was seen in few cases. Conclusion: Pleural effusion is a rare finding in SARS COV-2 induced bronchopneumonia. Effusion is seen with a high CORADS score, and they carry a high risk of mortality. This study suggests having SARS CoV-2 infection as a differential diagnostic entity when we find the mentioned morphological and biochemical findings as discussed in the present study.

Effect of different types of smoking on oral health and on cellular changes in oral mucosa

ObjectivesSmoking is known to be a major risk factor for several diseases. Recently, electronic cigarettes have been introduced to the market; however, their effect on oral health has not been well studied yet. We aim to compare the effect of different types of smoking on oral health and to evaluate cytomorphological changes in oral mucosa among different types of smokers. Study designA total of 112 participants were recruited, conventional cigarette smokers (n = 28), electronic cigarettes smokers (n = 26), hookah smokers (n = 29) and non-smokers (n = 29). Oral examination, brush cytology and salivary flow test were performed. Cytological smears were stained and examined for cytomorphological changes. Oral symptoms, type, and frequency of smoking were obtained through questionnaires. SPSS program was used for statistical analysis. ResultsMost of the oral symptoms reported in this study were among conventional and electronic cigarettes smokers. While hookah smokers reported the least oral symptoms. Increase in DMFT and gingival index were observed among all smokers, mainly among conventional cigarettes smokers. Conventional cigarette and hookah smokers were found to have the most significant cellular changes. Electronic cigarette smokers had certain cellular changes as well. ConclusionAll types of smoking adversely affect oral health and can induce cellular changes in the oral mucosa.

Cytological Features of Oral Cytobrush Smears in Type II Diabetes Mellitus Patients

Oral cytology is a renewed field that aids in diagnosis and observation of possible epithelial changes associated with oral mucosal diseases. Aim; to study the main cytomorphological alteration in gingival and buccal smears from type II diabetics in relation to their hyperglycemic status. The study includes 40 type II diabetic patients (20 new-diagnosed and 20 treated diabetics patients) and 20 healthy persons of both sex. Papanicolaou stained smear were prepared from their cheek and gingiva. The morphological features of 100 unfolded epithelial cells were evaluated under light microscope. Results of this study show that diabetics’ oral mucosa cells characterized by large nuclei with frequent evidence of binucleation, granular chromatin, prominent nucleoli. However, there was frequent small blue cytoplasm and buccal smears showed altered keratinization. As conclusion oral cytology from type II diabetics is associated with detectable cytomorphological changes that is site specific and indicate epithelial cell regeneration and degeneration with altered keratinization especially in buccal mucosa.

Influence of photoperiod and exogenous melatonin on testis morpho-physiology of sexually mature guinea fowl (Numida meleagris)

The biological effects of simulated photoperiod and melatonin on the control of reproduction of guinea fowls (Numida meleagris) are not well understood. Herein, thirty (30) sexually mature guinea fowl cocks were randomly assigned to 1–6 groups (n = 5) and subjected to different photoperiodic regimes in the presence or absence of exogenous melatonin (Mel; 1 mg/kgBW/day, i/m) for eight weeks. Testes of the euthanized cocks were processed for gross morphology, histological, histochemical, and oxidative stress markers. Testosterone concentration was determined in serum samples using the enzyme-linked immunosorbent assay (ELISA) technique. We observed an increase in testicular size in the Mel and Non-Mel groups under long-day (LD) photoperiods, and in the Non-Mel group under short-day (SD) photoperiod. Conversely, the testicular size was drastically reduced in the Mel group for SD. Seminiferous tubules in the Mel and Non-Mel groups of the SD showed cytomorphological changes, including degenerated cells, focal vacuolations, and depletion of germinal epithelium. However, the germinal epithelium appeared to be complete and active in both the Mel and Non-Mel groups for the LD. In all groups, the testes showed positive staining for PAS with varying intensities. There was a significant difference in PAS-staining intensity between different photoperiodic regimes and exogenous melatonin. The study observed the interaction between photoperiods and exogenous melatonin on glutathione reductase (GSH), malondialdehyde (MDA), and serum testosterone. Overall, the results indicated that a long-day (LD) photoperiod, combined with exogenous melatonin, enhanced reproductive activity in male guinea fowl by increasing testicular size and serum testosterone concentration.

THE EFFECT OF PARGILINE, SPERMINE AND THEIR COMBINATIONS ON ANDROGEN-DEPENDENT AND ANDROGEN-INDEPENDENT HUMAN PROSTATE CANCER CELLS in vitro

Summary. A series of studies on enhancing the cytotoxic effect of spermine on human prostate cells by affecting the enzymes of its metabolism has been continued. Aim: to investigate effects of pargyline, spermine and their combinations the survival, ζ-potential and surface charge and cytomorphological properties of human prostate cancer cells in culture. Object and methods: studies were conducted on cell cultures: differentiated androgen-dependent LNCaP cell line and low-differentiated androgen-independent DU-145 cell line. Cell survival was determined in the trypan blue assay. The effect of the investigated compounds, both individually and in combination, on the viability and proliferative activity of cells was assessed by the colorimetric method – by staining living cells with a solution of crystal violet in methanol. Biophysical parameters (ζ-potential and surface electric charge density) were determined according to the method cellular microelectrophoresis. Cytomorphological characteristics were studied on preparations of cells grown on coverslips, fixed and stained with hematoxylin and eosin (magnification 1000×). Results: for LNCaP cells, the IC50 of pargyline was 2.2 mM, and the IC50 of spermine was 1.5 mM. For DU-145 cells, the IC50 of pargyline was 2.35 mM, the IC50 of spermine was 5.0 mM. According to biophysical indicators, spermine caused an inversion of the sign of the electric charge in cells of the LNCaP line. Pargyline did not exert such an effect, the charge remained negative, demonstrating an inverse dependence on its concentration. The combination of pargyline and spermine increased the cytotoxic effect of spermine on LNCaP cells, but did not affect the value of their ζ-potential and surface charge density, the positive sign of which, caused by the effect of spermine, was preserved. At the same time, pargyline completely eliminated spermine-induced inversion of the sign of the total surface charge of DU-145 cells, returning it to the normal negative values. Conclusions: it is shown that the differentiated androgen-dependent cells of the LNCaP line with the combined use of pargyline and spermine demonstrated an enhancement of the cytotoxic effect of spermine. At the same time, low-differentiated cells of the DU-145 line, which do not have androgen receptors, were more resistant to the effects of both pargyline and pargyline in combination with spermine. Pargyline also reversed spermine-induced inversion of the sign of the total surface charge of DU-145 cells, returning it to normal negative values. Cytomorphological changes that occurred under the influence of both pargyline alone and in its combination with spermine demonstrate mainly the apoptotic pathway of cell death in the androgen-independent DU-145 line, while in the androgen-dependent LNCaP line, a smaller proportion of cells were in a state of apoptosis.

Environment Friendly Rehydrants for Air-Dried Oral Buccal Smears - A Comparative Study with Conventional Method.

Cytopathology is defined as a branch of pathology which involves studying diseases and their diagnosis. The conventional method involves fixing the cells in alcohol, but its poor availability in ill-equipped areas can pose problems. One solution to this problem is air-drying of smears followed by rehydration. In this study, we intend to assess the cytomorphologic changes in air-dried smears rehydrated using various agents and to correlate them with the smears prepared using conventional wet technique. Also, we attempt to find the alternative to alcohol which can be used in less equipped areas. Three smears each were prepared from 24 patients (12 healthy and 12 with habits). A total of 72 smears were prepared. One smear from each patient was used as control group (alcohol fixation - total number: 24 smears). The remaining two smears were air-dried and rehydrated and were taken as the test group (test group A - rehydrated with aq. glycerine; total number: 24 smears, test group B - rehydrated with coconut oil; total number: 24 smears) for 5-10 minutes. Then, all 72 slides were stained by standard Papanicolaou stain. The slides were examined for cytomorphologic changes and a score was assigned from 0 to 3. Our study proved aq. glycerine to be a superior rehydrant in comparison to conventional wet fixed smears or coconut oil. Also, in studies where cytoplasmic evaluation is required, coconut oil can be used due to its superior quality.

Understanding cytomorphologic changes in Pap tests of transgender men on testosterone therapy

Abstract Introduction/Objective Pap test findings in transgender men on testosterone therapy can often lead to misdiagnosis of dysplasia, especially in the setting of a positive HPV test. We reviewed relevant literature to analyze the cytomorphological nuances and applicable correlations to subsequent pathology in Pap tests of transgender men on testosterone therapy. Methods/Case Report We searched peer-reviewed journals on PubMed, synthesizing five articles published from 2018-2022 that specifically address cytomorphological findings on cervical Pap tests and biopsy results in transgender men on testosterone therapy. Results (if a Case Study enter NA) Various cytological changes, such as small parabasal-type cells and transitional cell metaplasia (TCM) can be mistaken for dysplasia in transgender men. In fact, these changes are representative of atrophy due to testosterone, as such findings have been observed in patients undergoing tamoxifen therapy. Small parabasal-type cells are described as clusters or sheets with intermittent nuclear enlargement and hyperchromatic nuclei, whereas TCM comprises flat sheets of cells with oval to spindle-shaped nuclei, powdery chromatin, and prominent nuclear grooves. Relevant clinical history can often help clarify cytologic diagnoses, as Pap tests that show atrophic changes can be misinterpreted as being abnormal. In several cases described in the literature, original diagnoses of atypical squamous cells of undetermined significance (ASCUS) were re-interpreted as atrophic changes after eliciting a history relevant for testosterone therapy. Small parabasal-type cells found on cytology have correlated with sections of the ectocervix that have groups of small hyperchromatic cells attached to the surface of atrophic cervical epithelium or areas of TCM. These cells may be mistaken for endometrial cells, especially in patients over 45 years old. Distinguishing atrophic changes from dysplasia on surgical pathology may modify cytologic diagnoses. Conclusion Transgender men on testosterone therapy often retain their cervix, necessitating ongoing cervical cancer screening. However, Pap test results from these individuals often exhibit atrophic changes through the presence of small parabasal-type cells and TCM. Differentiating between atrophy and dysplasia can pose a diagnostic challenge, especially in the setting of positive HPV testing or inflammation. Continued analyses into these cytomorphological nuances will assist providers in refining their diagnostic approaches and providing personalized care to transgender men.

Mitochondrial dsRNA from B-Precursor Acute Lymphoblastic Leukemia Cells Induce Formation of Cancer Associated Fibroblast in the Bone Marrow Leading to Formation of a Chemoprotective Bone Marrow Niche

Precursor B-cell Acute Lymphoblastic Leukemia (B-ALL) is typically a highly chemosensitive malignancy with the majority of children being cured by chemotherapy alone. Adult ALL is often chemosensitive at initial treatment, but long term disease free survival is less common. It is widely accepted that outcome in ALL closely relates to underlying genetic lesion (Moorman, Hematologica 2016) but no specific mechanisms by which ALL genetics influence outcome have been identified. We have previously demonstrated that an ‘activated mesenchymal stromal cell (MSC) niche’ develops in response to reactive oxygen species (ROS) -inducing chemotherapy in which mitochondria are actively transferred along tunnelling nanotubes to ‘rescue’ B-ALL cells from chemotherapy (Burt, et al Blood 2019). According to recent evidence that chemotherapy resistant ALL sub-clones may exist at diagnosis (Dobson et al Cancer Discovery 2020), we hypothesised that B-ALL cells may themselves directly activate bone marrow stromal cells to generate cancer associated fibroblasts (CAF) from MSC. To test this hypothesis, we first used our previously described in vitro niche model wherein either the HS27a MSC cell line or healthy donor (HD) MSCs are co-cultured with either B-ALL cell lines or primary patient B-ALL cells. Three of five B-ALL cell lines and 3 of 4 primary B-ALL samples tested, induced activation of the MSCs, as characterised by cytomorphological changes identified by confocal microscopy, up-regulation of CAF-relevant gene expression and increased IL6, IL8 and CCL2 protein secretion. The ability of the B-ALL cell lines to induce CAF correlated directly with the intrinsic ROS levels of the cell as determined by CellROX assay. In vivo, the CAF-inducing ALL cell lines also activated murine MSCs when established as xenografts in NOD-SCID gamma (NSG) mice. Immunohistochemistry of NSG murine femora revealed a very significant expansion in nestin+ cells after exposure to CAF-induing ALL cells compared to non-CAF-inducing controls. We established that the ALL cells activated MSC without contact, both from exposure of MSC to ALL cells in transwell and from exposure to B-ALL cell conditioned media (CM). RNA sequencing of HS27a stromal cells exposed to CAF-inducing B-ALL cell CM compared with non-CAF inducing controls unexpectedly showed that the most highly upregulated pathways were those involved in RNA sensing. Using a smaller gene panel selected from among the RNA-sensing pathway genes, we confirmed that this was also the case for primary, HD MSCs upon exposure to primary ALL patient samples. As a result, we sought evidence for the role of mitochondrial dsRNA, which has been shown to activate the anti-viral signalling pathway (Dhir et al, Nature 2018). Using the J2 anti-dsRNA monoclonal antibody in both confocal and flow cytometric assays, we observed very significantly higher levels of dsRNA in MSC which had been exposed to CAF inducing B-ALL cell CM than in non-CAF inducing B-ALL or MSC alone controls. We confirmed by RT-PCR for mitochondrial genes on dsRNA immunoprecipitated from CAF inducing B-ALL cell CM that the dsRNA was of mitochondrial origin. Using ELISA, we quantified dsRNA from total cell RNA extracted from paired primary B-ALL patient samples at diagnosis and in remission. We found significantly higher levels of dsRNA at diagnosis than in remission, with it being undetectable in most remission samples. Moreover, we confirmed by flow cytometry the presence of dsRNA in 6 B-ALL patient derived xenografts which was absent in the control NSG mice without leukemia. We used DMSO to degrade dsRNA from CAF inducing B-ALL cell CM, as is often done in Sanger sequencing and confirmed the degradation by ELISA. We then quantified gene expression of a selection of stromal cell activation marker genes, as well as genes in the RNA sensing pathway after exposure to either DMSO-treated B-ALL cell CM or non-treated control. There was a 2 to 4-fold reduction in expression of IL6, IL8 as well as of RIG-I, MX2, IFIH1, MAVS and OAS1 expression in DMSO-treated condition compared to control. We confirmed that IL6, IL8 and CCL2 protein secretion was also significantly abrogated in the test but not the control condition. We propose a model in which ROS-induced mitochondrial dysfunction in ALL cells leads to release of dsRNA which in turn activates bone marrow stromal cells leading to protective niche formation, despite lack of prior exposure to cytotoxic agents.

Dynamics of indicators of the functional state of the body and cytomorphological changes in wrestlers during the performance at the championship of Ukraine in Ukrainian belt wrestling

The issue of the functional state of the body of Ukrainian belt wrestlers is poorly studied. There are isolated research data on the improvement of the functional state of the body of wrestlers as a result of training. Studying the issues of the functional state of the body of wrestlers of various types of wrestling is extremely important, as it determines the athlete's success during competitions and the growth of skill during many years of sports training. The purpose of this work was to determine the physiological features of the response of the body of Ukrainian belt wrestlers during the Championship of Ukraine on April 29, 2023.
 As a result of the research, we established that in the pre-start period, the indicators of the cardiovascular system and respiratory system functioning of all examined wrestlers correspond to the physiological state of pre-start readiness of the athlete's body with a predominance of sympathetic vegetative influences. The functional indicators of the wrestlers after the termination of the performance at the Championship of Ukraine for Ukrainian belt wrestling differ depending on the number of matches they have held and the intensity of the physical load. The winner of the competition, having held 4 winning matches without a break between them, was subjected to physical stress at the level of submaximal power. The winners, who played a smaller number of matches with breaks in between, managed to restore the functional state of the cardiovascular system and respiratory system without experiencing such a load. The functional indicators of wrestlers who lost matches and stopped participating in competitions indicate an insufficient functional reserve of the cardiovascular system and respiratory system. The changes in the cytomorphological indicators of the buccal epithelium that we obtained indicate a significant strain on the body's functional systems, signs of dehydration in the winner who played four matches. In other athletes, there are certain signs of discomfort and strain on the body's functional systems (cytolysis, karyolysis, karyorrhexis, the presence of binucleated cells and leukocytes).

Quantitative Analysis of Bone Marrow Features Highlights Heterogeneity in Myelofibrosis Patients Treated with Zinpentraxin Alfa in a Phase II Clinical Study

Quantitative image analysis has potential to transform the interpretation of bone marrow trephine (BMT) samples in myeloproliferative neoplasms (MPN) and improve the evaluation of anti-fibrotic therapies. To investigate the utility of recently developed algorithms evaluating reticulin fibrosis and megakaryocyte features in myelofibrosis (MF), we analysed samples from a multi-center, phase II study of zinpentraxin alfa (ZPN, PRM-151). ZPN is a recombinant form of human pentraxin-2 (PTX2) that has shown clinical activity as monotherapy and in combination with ruxolitinib (RUX) in a phase II trial in patients with Int-1/-2 or high risk MF (NCT01981850). In stage 2 of this study, patients ineligible for, intolerant of, or with an inadequate response to RUX were randomised to receive 0.3, 3.0, or 10.0 mg/kg ZPN on Days 1, 3, and 5 of Cycle 1, and every 4 weeks thereafter for up to 9 cycles. We demonstrate the potential of quantitative image analysis to augment and refine conventional expert histological assessment of MF. Reticulin and H+E-stained BMTs from three timepoints (screening, C4D1 and C9D29) were analysed from 50/97 patients enrolled. Manual assessment (MA) of marrow fibrosis and megakaryocyte features was performed by blinded, independent central review. Assessment of fibrosis using the Continuous Indexing of Fibrosis (CIF) score and analysis of megakaryocytes was performed by automated analyses ( Ryou H, Leukaemia, 2022). There was a moderate correlation between the automated average CIF score and manual fibrosis grade (Spearman's rho = 0.39). However, there was significant overlap in the distribution of CIF scores between samples manually assigned to MF-2 and MF-3. Approximately 38% of MF-2 samples fell within the interquartile range of CIF distribution observed in MF-3, and around 48% of MF-3 samples fell within the interquartile range observed in MF-2. Next, we visualised the screening BMTs in fibrosis PCA (principal component analysis) space to capture both average CIF score and Shannon entropy (‘unevenness‘) of CIF score for each sample. This revealed significant heterogeneity in reticulin fibrosis at screening when compared to an independent reference cohort of 130 MPN samples ( Ryou H, Leukaemia, 2022), likely reflecting the effects of pre-treatment [ Figure 1]. To track fibrosis across all timepoints we plotted the average CIF score for each sample. This revealed striking variability in fibrotic response within the trial cohort, with 15 of 42 patients (36%) demonstrating improvement in average CIF score [ Figure 2]. Notably, improvements in average CIF score by cycle 9 appeared to be most marked in patients with higher CIF scores at screening. No significant ZPN dose-dependent effect on fibrosis was observed. We next measured megakaryocyte density (megakaryocytes per unit area of intertrabecular space) and compared it to MA. We found a strong correlation (Spearman's rho = 0.81) between automated and MA of megakaryocyte density, with no significant change across any of the treatment arms. For megakaryocyte clustering (defined as ≥ 3 megakaryocytes in direct contact) we observed a moderate to strong correlation (Spearman's rho = 0.62) between MA and automated assessment, with no significant change across any of the treatment arms. Finally, we sought to compare the sample megakaryocyte cell features (cytomorphology and topology) at screening with those of an independent reference cohort of 88 MPN samples ( Sirinukunwattana, Blood Adv., 2020). Strikingly, not only was there marked heterogeneity in megakaryocyte features between patient samples at screening, but the observed features were significantly different to those of newly diagnosed MF. No significant changes in megakaryocyte cytomorphology or topology were seen across any of the treatment arms. In summary, our findings reveal considerable BMT morphological heterogeneity in patients participating in a Phase II trial of ZPN. This likely reflects poorly understood and under-recognised variation in morphological features encountered in patients with longstanding and/or pre-treated MF when compared to newly diagnosed patients. This highlights the potential of such variability to confound the evaluation of novel therapeutics in MPN, and emphasises the utility of robust quantitative methods to analyse and visualise morphological features in clinical study samples that can complement conventional manual assessment.

Cord Blood as a Corrector of Hematological and Cytomorphological Characteristics of Lymphohemopoietic Organs in Experimental Atopic Dermatitis

Background. Atopic dermatitis (AD) is an inflammatory skin disease caused by a shift in the interaction between cells of the immune system and the skin. Steroid anti-inflammatory drugs used to treat AD often lead to numerous side effects. Therefore, the development of new natural multifunctional preparations with immunomodulatory activity is of great importance. Objective. To determine the changes in indices of the leukocytes cluster of peripheral blood and determine the peculiarities of cytomorphological changes in the spleen and lymph nodes (LN) of rats with induced AD before and after administration of cryopreserved (cHCBL) and lyophilized (lHCBL) forms of human cord blood leukoconcentrate. Methods. The experiments were conducted using 6-month-old Wistar rats. AD was induced by topically applying a 5% alcohol-acetone solution of dinitrochlorobenzene (DNCB) to the skin of the rats' back for 21 days. Each form of the preparation (cHCBL and lHCBL) was administered intraperitoneally in a 0.5 ml dose containing 5´106 cells, one day after the final DNCB treatment. The following parameters were assessed in the blood: leukocyte count and the neutrophil-to-lymphocyte ratio (N/L index). Сytomorphological characteristics were determined in the spleen and inguinal LN. Results. In AD, specific changes in the cytomorphological characteristics of the lymphohematopoietic complex (LHС) structures, namely the spleen and LN, were observed alongside alterations in the leukocyte cluster indices. The most notable changes in the spleen included the dominance of the white pulp over the red pulp and the prevalence of follicles of stages II and III of development in the LN. Animals with AD displayed pronounced leukocytosis and an increased N/L index in their blood. The study demonstrates the positive corrective effects of cHCBL and lHCBL on the morphological characteristics of LHC and the specified blood parameters. Conclusions. The effectiveness of the cHCBL and lHCBL use for correcting indices of leukocytes cluster of peripheral blood and the structural organization of lymphohemopoietic organs in the development of AD have been proven.

The kynurenic acid analog SZR104 induces cytomorphological changes associated with the anti-inflammatory phenotype in cultured microglia

We previously showed the anti-inflammatory effects of kynurenic acid (KYNA) and its brain-penetrable analog N-(2-(dimethylamino)ethyl)-3-(morpholinomethyl)-4-hydroxyquinoline-2-carboxamide (SZR104) both in vivo and in vitro. Here, we identified the cytomorphological effects of KYNA and SZR104 in secondary microglial cultures established from newborn rat forebrains. We quantitatively analyzed selected morphological aspects of microglia in control (unchallenged), lipopolysaccharide (LPS)-treated (challenged), KYNA- or SZR104-treated, and LPS + KYNA or LPS + SZR104-treated cultures. Multicolor immunofluorescence labeling followed by morphometric analysis (area, perimeter, transformation index, lacunarity, density, span ratio, maximum span across the convex hull, hull circularity, hull area, hull perimeter, max/min radii, mean radius, diameter of bounding circle, fractal dimension, roughness, circularity) on binary (digital) silhouettes of the microglia revealed their morphological plasticity under experimental conditions. SZR104 and, to a lesser degree, KYNA inhibited proinflammatory phenotypic changes. For example, SZR104 treatment resulted in hypertrophied microglia characterized by a swollen cell body, enlarged perimeter, increased transformation index/decreased circularity, increased convex hull values (area, perimeter, mean radius, maximum span, diameter of the bounding circle and hull circularity), altered box-counting parameters (such as fractal dimension), and increased roughness/decreased density. Taken together, analysis of cytomorphological features could contribute to the characterization of the anti-inflammatory activity of SZR104 on cultured microglia.

Urine cytology in patients with gender confirmation surgery and hormone therapy: evaluation of urine cytology performance in an underserved patient population.

There is a practice gap and educational need regarding urine cytology (UC) performance in patients with history of gender confirmation surgery (GCS) and/or hormone therapy (HT). This potentially impacts diagnostic accuracy in this medically underserved population. We report a methodology that identifies relevant cases and evaluates the performance of UC in this cohort. Two institutional pathology archives from 2000 to 2021 were searched using relevant keywords to identify UC specimens from patients with GCS and/or HT for this retrospective study. For each specimen, patient demographics, relevant clinical history, and history of HT and/or GCS were noted. Each case was blindly reviewed by a cytopathologist according to The Paris System. A total of 32 UC specimens from 15 patients with history of GCS and/or HT were identified. There were 13 male to female and 2 female to male transgender patients. The original diagnosis was negative for high-grade urothelial carcinoma (NHGUC) in 24 of 32 (75%) and atypical urothelial cells (AUC) in 8 of 32 (25%) cases. The most common atypical features were irregular nuclear membranes and prominent small nucleoli in 7 of 8 (87.5%). Degenerative changes were present in 5 of 8 (62.5%). On re-review, with relevant clinical history, 100% of cases were re-classified as NHGUC. The original diagnosis of AUC in these cases likely reflects reactive changes post GCS and/or HT. This cohort may be at risk of AUC overdiagnosis, particularly if the pathologist is unaware of this clinical history. Pathologists need to recognize reactive cytomorphologic changes in these patients. Further multi-institutional studies are warranted to expand knowledge about UC performance in these patients.

Цитоморфологічні та цитогенетичні характеристики букального епітелію осіб, які мешкають в умовах «хворої будівлі»

The micronucleus test on buccal cells is a sensitive, minimally invasive and quite common method of cytogenetic studies. Aim. Study of cytomorphological and cytogenetic characteristics of the buccal epithelium using the micronucleus test in persons exposed to volatile organic compounds in the conditions of a "sick building". Materials and methods. The condition of the buccal epithelium was studied in 237 residents of apartment buildings and offices who complained of an unpleasant smell in the premises (male – 117, female – 120). Buccal cells were stained with 2.5 % acetoorcein solution and 1 % light green. Microscopy of the preparations was carried out using an optical microscope OPTON Axioskop (Germany) with magnification by oil immersion – x1000. Morphometric analysis of cell parameters and calculation of the percentage ratio of different forms of epitheliocytes was carried out using the Excel program. Calculations of the frequency of formation of micronuclei were carried out according to the order of the Ministry of Health of Ukraine dated 13.03.2007 № 116 "On the approval of methodological recommendations: Survey and zoning of the territory according to the degree of influence of anthropogenic factors on the state of environmental objects using cytogenetic methods." Statistical data processing was performed using the IBM SPSS Statistics 29.0.0.0 program (Student's t-test; Mann-Whitney; ANOVA: Tukey; T3-Dunnett), at p ≤ 0.05. Results. Cytomorphological and cytogenetic changes in buccal cells were detected in a wide range of frequencies of 0.3–7.0 ‰; normal cells – 81 %; karyorrhexis and mast cells – 5 %, nuclear doubling and vacuolar dystrophy – 2 %, phagocytosis-apoptosis and absence of a nucleus – 1 %, colonization by microflora – 1 %, micropoison – 0.6 %, multinucleated cells – 0.5 %, protrusion of the nucleus "broken "egg" – 0.5 %, "tongue" nucleus protrusion – 0.3 %. According to the MN-test, it was established that a strong correlation between the parameters of the frequency of MN formation and increasing age (R2 = 0.860) was characteristic of females, however, the degree of correlation was weak among males (R2 = 0.495). It was also established that the maximum rate of formation of micronucleus was observed among males in the age groups of 15–39 years and 65 years and older, and among females – in the age group of 65 years and older. Conclusions. In general, studies confirm that buccal epitheliocytes are sensitive to various exogenous and endogenous factors, which affects the cytomorphological and cytogenetic features of buccal cells, and these features have a rather wide spectrum. Keywords: indoor air, "sick building syndrome", cytogenetic studies, buccal epithelium, micronucleus test.