Nuclear Diameter Research Articles

Abstract A017: High-throughput microfluidic enrichment of circulating tumor cells from entire diagnostic leukapheresis for comprehensive liquid biopsy

Abstract Circulating Tumor Cells (CTCs) in blood containing a complete set of single-cell analytes, including high-molecular-weight DNA, intact RNA, and both intracellular and cell-surface proteins serve as a promising tool in cancer diagnosis and prognosis through liquid biopsy. However, their clinical utility has been limited by their low blood concentration and inadequate enrichment technology. Diagnostic leukapheresis holds promise for isolating sufficient CTCs, but enriching ultra-rare cells from large, complex fluid volumes remains challenging. To address this, we developed the LPCTC-iChip, a high-throughput microfluidic device that uses high-flow channels and force-amplifying magnetic lenses to deplete hematopoietic cells, allowing tumor epitope-agnostic screening of massive blood volumes. Here, we present the first application of LPCTC-iChip to entire diagnostic leukapheresis products (leukopak) from seven patients with metastatic cancer, processing an average of 5.83 liters of blood per patient. Enumeration and morphology characterization of CTCs using multispectral immunofluorescence imaging identified high CTC yields - a mean of 10,057 CTCs per leukopak and showed notable intra- patient heterogeneity. Nuclear and cell diameter of CTC varies within individual patients, with 67% overlapping in size with patient-matched white blood cells. Paired single-cell DNA and RNA sequencing uncovered subclonal aneuploidy and distinct signaling pathways within CTCs with shared, clonal copy number variation (CNV) patterns. A subset of small aneuploid cells lacking epithelial markers was enriched for neuroendocrine characteristics in prostate cancer samples. The mutations found by FDA-approved genetic tests in either biopsied metastatic lesions or blood-based ctDNA sampling were reproduced by whole exome sequencing of a pool of CNV-confirmed CTCs. In addition, the sequencing revealed additional cancer-related high allele frequency variants that are not detectable with ctDNA or tissue needle biopsies. Overall, this high-throughput CTC enrichment from complete leukapheresis combined with multispectral imaging and single cell sequencing technology enables a comprehensive, cell-based liquid biopsy approach for early cancer detection and therapeutic responses monitoring. Citation Format: Avanish Mishra, Shih-Bo Huang, Taronish Dubash, Risa Burr, Jon F. Edd, Ben S. Wittner, Quinn E. Cunneely, Victor R. Putaturo, Akansha Deshpande, Ezgi Antmen, Kaustav A. Gopinathan, Keisuke Otani, Yoshiyuki Miyazawa, Ji Eun Kwak, Sara Y. Guay, Justin P. Kelly, John Walsh, Linda T. Nieman, Isabella Galler, PuiYee Chan, Michael S. Lawrence, Ryan J. Sullivan, Aditya Bardia, Douglas S. Micalizzi, Lecia V. Sequist, Richard J. Lee, Joseph W. Franses, David T. Ting, Patricia A. R. Brunker, Shyamala Maheswaran, David T. Miyamoto, Daniel A. Haber, Mehmet Toner. High-throughput microfluidic enrichment of circulating tumor cells from entire diagnostic leukapheresis for comprehensive liquid biopsy [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A017.

Morphological Changes Of The Heart In Sudden Death Psychiatric Patients

Although sudden cardiac death is often reported in epileptic or psychiatric patients, the etiologyis not fully understood. The aim of this study was to examine whether morphological changes of the heart contribute to sudden cardiac death in epileptic or psychiatric patients, compared with control decedents.Cases of sudden cardiac deathsdue to fatal arrhythmia in patients with epilepsy or psychiatric disease were selected from forensic autopsies performed at the Shiga Prefecture from 2014 to 2021.As the control group, autopsied healthy decedents were selected. The heart weight to normal heart weight ratio, ventricular wall thickness, cardiomyocyte width, nuclear diameter and nuclear density of the left and right ventricle and interventricular septum were examined. The left and right ventricular wall thickness and cardiomyocyte to be smaller in epileptic patients, however, no significant differences were found. The nuclear diameter/body surface area of the left ventricle was significantly smaller in epileptic and psychiatric patients than in control patients.The smaller nuclei of cardiomyocytes in epileptic and psychiatric patients may relate to the etiology of sudden cardiac death.

Alcoholic Hepatitis: A Paradigm shift of Prevention and Treatment Analyzed at Cellular level by Histo/Pathological Micrometric Assessment.

Objective: To evaluate role of Vitamin C and Naproxen in Alcohol-induced liver toxicity by micrometric technique (hepatocyte count, size and nuclear diameter) in albino rats.Methodology: This experimental interventional study was conducted at the sheikh Zayed medical college and hospital Rahim yar khan from December 2022 to March 2023. 60 albino rats with equal gender distribution were selected based on the random probability sampling technique and placed in three groups of 20, again with an equal gender distribution (1:1). Group A (positive control) was given purified Ethanol for 10 days., Group B was given Vitamin C and NaproxenProphylaxis for 7 days and later intoxicated with Ethanol for 10 days and Group C was simultaneously Administered Vitamin C, Naproxen, and Ethanol for 10 days. The hepatocyte count, size and nuclear diameter of all the groups were recorded by micrometric technique using ocular and stage micrometric scales. Data analysis was conducted using SPSS 24.0 and P-Value of ≤0.05 considered Statistically Significant.Results: Group A animals experienced decreased number of hepatocyte count, increase in size of cell and decrease in nucleus size; Group B showed similar picture as group “A” but less in intensity and Group C showed nearly normal hepatic architecture with hepatocyte count, size and nuclear size close to normal ranges.Conclusion: The anti-oxidative and anti-inflammatory effects of Vitamin C and Naproxen demonstrated significant hepatoprotective effects on liver histologic architecture, providing safety from alcohol-induced liver injury. Keywords: Naproxen, Prophylaxis, liver toxicity, alcoholic liver disease, vitamin C.

Nuclear deformation and dynamics of migrating cells in 3D confinement reveal adaptation of pulling and pushing forces.

Eukaryotic cells show an astounding ability to remodel their shape and cytoskeleton and to migrate through pores and constrictions smaller than their nuclear diameter. However, the relation of nuclear deformation and migration dynamics in confinement remains unclear. Here, we study the mechanics and dynamics of mesenchymal cancer cell nuclei transitioning through three-dimensional compliant hydrogel channels. We find a biphasic dependence of migration speed and transition frequency on channel width, peaking at widths comparable to the nuclear diameter. Using confocal imaging and hydrogel bead displacement, we determine nuclear deformations and corresponding forces during confined migration. The nucleus deforms reversibly with a reduction in volume during confinement. With decreasing channel width, the nuclear shape during transmigration changes biphasically, concomitant with the transitioning dynamics. Our proposed physical model explains the observed nuclear shapes and transitioning dynamics in terms of the cytoskeletal force generation adapting from purely pulling-based to a combined pulling- and pushing-based mechanism with increasing nuclear confinement.

Cytological and Cytometric Analysis of Epithelial Cell Changes Under the Surface of Acrylate Prosthesis in Diabetic Patients

Background: The aim of this study was to assess cytological alterations of the squamous epithelium of the alveolar ridge mucosal surface under the acrylate prosthesis in T2D patients. Methods and Results: The subjects of interest were patients in whom the total acrylate prosthesis had been applied 3-5 years or more prior to the examination. The subjects were divided into two groups: 30 adult subjects with T2D (T2D group) and 30 adult subjects without T2D (control group). Both groups were over 49 years of age. Cytological smears were obtained by cytobrush from the mucosal surface of the gingival crest underlying the acrylate prosthesis. The following parameters were assessed: type of cells (basal, intermediate, superficial, superficial without nucleus, and parakeratotic), proportions of the types of cells, cytoplasmic diameter, nuclear diameter, and nucleus-to-cytoplasm ratio. An independent sample t-test was used to compare the two study groups. The percentage of superficial cells was significantly lower in the T2D group than in the control group (P=0.001). The T2D group tended to have a higher rate of parakeratotic cells (13.0±11.12 vs. 8.07±7.92, P=0.052). Superficial cells in the T2D group had a significantly lower mean cytoplasmic diameter than the control group (32.7947±8.61929 µ vs. 36.6383±4.32228 µ, P=0.03). Additionally, the nucleus-to-cytoplasm ratio of intermediate and superficial cells in the T2D group was significantly higher than in the control group (0.2407±0.07206 vs. 0.2000±o.03291, P=0.007 and 0.2573±0.06330 vs. 0.2280±0.03178, P=0.027). Conclusion: The results of our study show that total acrylate prostheses in diabetic patients are responsible for the disrupted maturation of squamous epithelial cells. This is reflected in smaller superficial cells, increased parakeratosis, and the higher nucleus-to-cytoplasm ratio of intermediate and superficial cells.

Nuclear long diameter as a new criterion to distinguish high-grade urothelial carcinoma cells from benign reactive cells.

Recently, the nuclear area has attracted attention as a morphological parameter to differentiate high-grade urothelial carcinoma (HGUC) cells from benign reactive cells. The nuclear long diameter (NLD) strongly correlates with the nuclear area and is easy to subjectively estimate. Therefore, this study examined the usefulness of the NLD-to-neutrophil diameter ratio for detecting HGUC cells in urine cytology. This study included 29, 26 and 18 patients with HGUC, glomerular disease and urolithiasis respectively. An image analysis system was used to measure the NLD of HGUC and benign reactive cells (reactive renal tubular cells and reactive urothelial cells) and the neutrophil diameter that appeared in the voided urine in these cases. The NLD index was calculated using the NLD-to-neutrophil diameter ratio. We subsequently compared HGUC and benign reactive cells with respect to NLD and NLD indices. In addition, the HGUC cell group and benign reactive cell group were compared by selecting the five cells with the largest NLD and NLD index on each slide. The NLD and NLD indices of HGUC cells were significantly higher than those of benign reactive cells in all cells and in the five cells with the largest NLD and NLD indices. The cut-off value of the NLD index for detecting HGUC cells was 1.25 in all cells and 1.80 in the five cells with the largest NLD index. The NLD index is a useful parameter that can be introduced into routine microscopic examinations to differentiate HGUC cells from benign reactive cells.

Hepatoprotective Effect of Aqueous Neem Leaves Extract on Hepatocytes and Nuclear Diameter of Albino Wistar Rats Induced with Erythromycin Toxicity

Objective: To evaluate the hepatoprotective effects of Neem leaves extract on hepatocyte and nucleardiameters of Albino Wistar rats induced by erythromycin hepatotoxicity.Study Design: Laboratory-based Experimental study.Place and Duration of Study: The study was carried out at the Department of Anatomy and Animal House ofBaqai Medical University (BMU) in Karachi, Pakistan from January 2019 to March 2019.Methods: A total of 80 Albino Wistar rats were divided equally into 4 groups. The control group was markedas Group A with no interventions. In Group B only erythromycin was administered. For Group C botherythromycin and aqueous Neem leave Extract were given and for Group D only aqueous neem leave wasgiven. Animals were sacrificed and the liver weight of Albino Wistar rats was measured.Results: It was observed that rats of experimental control group A noticed no significant change inhepatocytes13.19± 0.44μm and nuclear diameters 7.18±0. 28μm (Figure.1) as this group had no interventionreceived. In group B hepatocyte diameter markedly increased by 16.54±0.6μm and a decrease in nucleardiameter of 5.20±0.30μm (p<0.01) with loss of architecture, ballooning, and edematous changes (Figure.2, 3and 4) was observed as this group received only erythromycin while group C received both erythromycin andaqueous Neem leaf extract showed histological architecture disrupted to some extent and hepatocytes14.49±80μm and nuclear diameter 6.13±0.17μm were started to improve its normal sizes (Figure.5). Group Dobserved no significant changes in hepatocytes 13.0±0.52μm and nuclear diameters 7.14±0.30μm as thisgroup only received aqueous neem leave extract (Figure.6).Conclusion: According to the current study, Erythromycin exerts oxidative stress and Changes in hepatocytesand nuclear diameters were noticeably improved by aqueous neem leaf extract, which is a powerfulantioxidant. How to cite this: Jamshed N, Raza I, Razzaq M, Jamshed N, Rasheed A, Faisal L. Hepatoprotective Effect of Aqueous Neem Leaves Extract on Hepatocytes and Nuclear Diameter of Albino Wistar Rats Induced with Erythromycin Toxicity. Life and Science. 2024; 5(2): 228-236. doi: http://doi.org/10.37185/LnS.1.1.572

ULTRAVIOLET RADIATION DOSIMETRY BY USING OF CN-85 , CR-39, LR-115 NUCLEAR TRACK DETECTORS

Ultraviolet radiation dosimetry was determined by using CN-85 , CR-39 ,LR-115 nuclear track detectors – NTD with measuring of ; number of track-NT , etching time-TB , nuclear track diameter - DT and etching velocity -VD . By this study appear the increasing in etching time-TB of NTD samples which irradiated by UV-radiation do not pure effect on the number of track - NT , comparing with un-irradiated samples . The increasing in UV-irradiation make decreasing in etching time- Topt for CR-39 , CN-85 , LR-115 nuclear track detectors with percent of 20% , 25% , 50% respectively , comparing with un– irradiated sample at the radiation dose 3.77x103 erg /mm2 . . The increasing of irradiation dose make increasing in nuclear track diameter-DT with increase in etching time-TB at the radiation dose 300x103erg / mm2 for LR-115 detector . This study showing there was increasing in the percent value of etching velocity -VB with increasing inradiation dose for CR-39 , CN-85 . The increasing in nuclear track diameter- DTwith increasing in radiation dose was appearing as a resulted of energy of radiationand producted free radicals which interact with chemical etching solution . Thisstudy optioned by using nuclear track detectors CN-85 , CR-39 for determinationthe radiation dosimetry through measuring of etching velocity -VB butter than LR-115 detector

Differentiating BRAF V600E- and RAS-like alterations in encapsulated follicular patterned tumors through histologic features: a validation study.

Differentiating BRAF V600E- and RAS-altered encapsulated follicular-patterned thyroid tumors based on morphology remains challenging. This study aimed to validate an 8-score scale nuclear scoring system and investigate the importance of nuclear pseudoinclusions (NPIs) in aiding this differentiation. A cohort of 44 encapsulated follicular-patterned tumors with varying degrees of nuclear atypia and confirmed BRAF V600E or RAS alterations was studied. Nuclear parameters (area, diameter, and optical density) were analyzed using a deep learning model. Twelve pathologists from eight Asian countries visually assessed 22 cases after excluding the cases with any papillae. Eight nuclear features were applied, yielding a semi-quantitative score from 0 to 24. A threshold score of 14 was used to distinguish between RAS- and BRAF V600E-altered tumors. BRAF V600E-altered tumors typically demonstrated higher nuclear scores and notable morphometric alterations. Specifically, the nuclear area and diameter were significantly larger, and nuclear optical density was much lower compared to RAS-altered tumors. Observer accuracy varied, with two pathologists correctly identifying genotype of all cases. Observers were categorized into proficiency groups, with the highest group maintaining consistent accuracy across both evaluation methods. The lower group showed a significant improvement in accuracy upon utilizing the 8-score scale nuclear scoring system, with notably increased sensitivity and negative predictive value in BRAF V600E tumor detection. BRAF V600E-altered tumors had higher median total nuclear scores. Detailed reevaluation revealed NPIs in all BRAF V600E-altered cases, but in only 2 of 14 RAS-altered cases. These results could significantly assist pathologists, particularly those not specializing in thyroid pathology, in making a more accurate diagnosis.

The prognostic role of single cell invasion and nuclear diameter in early oral tongue squamous cell carcinoma

BackgroundThe clinical significance of single cell invasion and large nuclear diameter is not well documented in early-stage oral tongue squamous cell carcinoma (OTSCC).MethodsWe used hematoxylin and eosin-stained sections to evaluate the presence of single cell invasion and large nuclei in a multicenter cohort of 311 cases treated for early-stage OTSCC.ResultsSingle cell invasion was associated in multivariable analysis with poor disease-specific survival (DSS) with a hazard ratio (HR) of 2.089 (95% CI 1.224–3.566, P = 0.007), as well as with disease-free survival (DFS) with a HR of 1.666 (95% CI 1.080–2.571, P = 0.021). Furthermore, large nuclei were associated with worse DSS (HR 2.070, 95% CI 1.216–3.523, P = 0.007) and with DFS in multivariable analysis (HR 1.645, 95% CI 1.067–2.538, P = 0.024).ConclusionSingle cell invasion and large nuclei can be utilized for classifying early OTSCC into risk groups.

Simulated microgravity improves maturation of cardiomyocytes derived from human induced pluripotent stem cells

Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) possess tremendous potential for basic research and translational application. However, these cells structurally and functionally resemble fetal cardiomyocytes, which is a major limitation of these cells. Microgravity can significantly alter cell behavior and function. Here we investigated the effect of simulated microgravity on hiPSC-CM maturation. Following culture under simulated microgravity in a random positioning machine for 7 days, 3D hiPSC-CMs had increased mitochondrial content as detected by a mitochondrial protein and mitochondrial DNA to nuclear DNA ratio. The cells also had increased mitochondrial membrane potential. Consistently, simulated microgravity increased mitochondrial respiration in 3D hiPSC-CMs, as indicated by higher levels of maximal respiration and ATP content, suggesting improved metabolic maturation in simulated microgravity cultures compared with cultures under normal gravity. Cells from simulated microgravity cultures also had improved Ca2+ transient parameters, a functional characteristic of more mature cardiomyocytes. In addition, these cells had improved structural properties associated with more mature cardiomyocytes, including increased sarcomere length, z-disc length, nuclear diameter, and nuclear eccentricity. These findings indicate that microgravity enhances the maturation of hiPSC-CMs at the structural, metabolic, and functional levels.

Cytomorphometric analysis: A possible adjunct for age estimation

Abstract Background: Age estimation plays a crucial role in identifying an unknown individual in the event of any mass disaster or crime. One of the important noninvasive procedures that may have an impact on determining an individual’s age is exfoliative cytology. Aim and Objective: Exfoliated buccal mucosa cells were used in the study to determine the participant’s age by assessing the changes in nuclear diameter (ND), nuclear–cytoplasmic (N: C) ratio, and cell diameter (CD). Materials and Methods: With the help of a moistened wooden spatula and a light scraping motion, buccal smears were taken from 100 healthy and normal individuals across various age groups. The samples were then preserved with 95% ethanol and stained using ultrafast Papanicolaou stain. Cell and nuclear diameter were measured for each of these cells using the ImageJ software, and N: C was calculated from the two parameters. The N: C was calculated from CD and ND for each subject. One-way ANOVA tests were used for statistical analysis. Results: A significant difference in the diameter of cell and nucleus with the individual’s age (P < 0.05) was noticed. The N: C ratio reduces with increasing age but is not consistent. Conclusion: Buccal smears show age-related alterations that could be utilized as an effective tool for age estimation.

Psychometric and cytomorphometric analysis of dental phobia in pediatric patients: Association between anxiety and cellular structure

ABSTRACT Background: Children frequently experience negative feelings, such as dental fear or anxiety, as a result of the stress of receiving dental care. Extremely anxious individuals resist therapy or completely avoid it, which exacerbates oral and other health issues. Aim: The goal of this study was to examine the cytomorphometric responses toward dental interventions and their potential interplay with psychometric indices of anxiety levels and previous dental experience, specifically anesthesia, and how they varied before and after the treatment in children. Materials and Methods: On informed consent, buccal mucosal samples were obtained from 20 children (33% males and females 67%) in the pediatric dentistry clinic before treatment and immediately posttreatment. Dental anxiety was assessed through specific questionnaires (Modified Child Dental Anxiety Scale faces version) completed by the children. Results: On the cellular level: Cellular diameter (CD), nuclear diameter (ND), nuclear area (NA), and cellular area (CA) were assessed (Magnus Pro software and hematoxylin and eosin staining) while scores from the questionnaire established anxiety levels. Compared to pretreatment, mean CD, CA, and NA were decreased following treatment, while ND was higher at the recall. Dental anxiety questionnaire scores were also significantly correlated with each other which turned out to be positive. Conclusion: As a result, in response to environmental inputs, mitochondrial cells dynamically alter in terms of morphology and functionality. Chronic structural and functional variation in mitochondria can result in functional adjustments which were correlated by cytomorphometric and psychological investigation.

Effects of acetylated distarch phosphate on the physicochemical characteristics and stability of the oyster sauce system.

In this study, the effect of different acetylated distarch phosphate (ADSP) ratios (0, 1%, 2%, 3%, 4%, 5%) on the physicochemical characteristics and stability of the oyster sauce (OS) system was investigated. The texture, water state, interactions, rheological properties, microstructure, and stability of OS samples were analyzed through the texture analyser, low-field nuclear magnetic resonance (LF-NMR), particle diameter and zeta potential, fourier-transform infrared spectroscopy (FTIR), rheometer, and microscopes. The results revealed that the addition of ADSP improved the firmness, consistency, cohesiveness, and water-holding capacity of OS. Moreover, ADSP reduced the average particle size and zeta potential of OS, indicating that electrostatic and steric stabilization existed in the ADSP-OS system. The addition of ADSP enhanced the hydrogen bonding and decreased water mobility for OS system, processing a more continuous and smooth structure. All ADSP-OS samples were typical non-Newtonian fluids with shear-thinning characteristics. In addition, the non-significant instability index changes of ADSP-OS over the whole storage period confirmed the excellent long-term stabilization capability of OS prepared with ADSP. This study provides a theoretical basis for starch-based sauce products and contributes to the development of sauce products.

Effect of season on histoarchitecture of pineal gland in buffalo (Bubalus bubalis).

The photoperiod and other seasonal variations are the key factors that affect reproduction and production of the animals. The pineal gland secretes melatonin hormone that affects several physiological functions of the body during different seasons. The present study was conducted to study the histoarchitectural and micrometrical changes in the pineal gland of buffalo (Bubalus bubalis) during different seasons of the year. Pineal glands of 30 adult female Jaffarabadi buffaloes were collected from the slaughterhouse during the winter, summer, and rainy seasons. Samples were processed by standard histological procedures and stained with various stains for histological and micrometrical observations. The pinealocytes constituted a major cellular portion of pineal parenchyma. The pinealocyte nuclei were lightly stained and more euchromatic during the winter season whereas darkly stained and slightly heterochromatic during summer. The calcium deposits occupied a larger area of pineal parenchyma during the summer as compared to the winter season. The pinealocyte density, the nuclear diameter of pinealocytes, and the number of argyrophilic nucleolar organizer regions (AgNOR) were highest during the winter season as compared to the summer and rainy seasons. The present study shows the influence of season on the histoarchitecture and histometry of the pineal gland of buffalo and indicated higher pineal activity during the winter season in this species.

Ethanol, Carnoy, and paraformaldehyde as fixative solutions for histological evaluation of preantral follicles in equine ovarian tissue

The most adequate fixative solution for equine ovarian tissue is still to be determined as a tool to evaluate the improvement of methodological studies in assisted reproductive techniques and fertility preservation. This study aimed to evaluate a short-time ethanol 70% (ST-EtOH, 45 min) exposure as an alternative fixative compared with two classically fixatives [Carnoy’s (CAR) solution and paraformaldehyde 4% (PFA)] at different fixation times (6 h, 12 h). The end points evaluated were morphology and classes of preantral follicles, follicular and stromal cell densities, and follicular and oocyte nuclear diameters in equine ovarian tissue. Ovaries (n = 6) from ovariectomized young mares were fragmented (3 × 3 × 1 mm; 20 fragments/ovary) and fixed in the tested treatments. Overall, a total of 11,661 preantral follicles were evaluated in 1444 histological slides. The ST-EtOH similarly preserved the preantral follicle morphometry and stromal cell density compared to the PFA fixative, regardless of the exposure time. Nonetheless, the CAR fixative solution had the greatest percentage of normal preantral follicles and the highest stromal cell density among all treatments. In conclusion, Carnoy’s solution must be preferred compared with ST-EtOH and PFA fixatives for studies concerning the cellular morphology of equine ovarian tissue. Moreover, ST-EtOH fixative is a good alternative for equine ovarian tissue when a quick histological evaluation is required instead of more time-consuming and expensive techniques. Additional studies concerning the impact of different fixatives on the ultrastructure of cellular populations and their compatibility with IHC and molecular techniques in equine ovarian tissue are warranted.

Investigating In Situ Expression of Neurotrophic Factors and Partner Proteins in Irreversible Pulpitis

IntroductionIn situ assessments of neurotrophic factors and their associated molecular partners have not been explored to date, particularly in humans. The present investigation aimed to explore the expressional dysregulation of neurotrophic factors (nerve growth factor [NGF], brain derived neurotrophic factor [BDNF], and NT4/5), their receptors (TrkA and TrkB), and their modulators (USP36 and Nedd4-2) directly in irreversibly inflamed human pulp tissues. MethodsForty samples each of healthy and irreversibly inflamed pulp were extirpated for the study. Immunohistochemical examinations were carried out for the anatomic changes and expression of neurotrophic factors and partner proteins. Expression was digitally quantified using the IHC profiler module of ImageJ and deduced as optical density. Statistical analyses were carried out by GraphPad Prism. ResultsDecrease in nuclear and vessel diameters was observed in irreversibly inflamed pulp tissues. NGF and BDNF were found to be significantly upregulated in symptomatic irreversible pulpitis (SIP), whereas no significant difference was observed in the expression of TrkA and TrkB. Expression of Nedd4-2, USP36, and TrkA was found positively correlated with the NGF in healthy pulp tissues. However, in SIP, positive correlation was only observed between the expression of USP36 and NGF. Among the ligands, BDNF expression was found positively correlated with NGF in healthy pulp but not with NT4/5. In the case of SIP, no correlation was observed between any neurotrophic factors. ConclusionsUpregulation of NGF, BDNF, USP36 and Nedd4-2 in SIP indicates dysregulation in the molecular events underlying the disease biology and could be exploited as potential markers for the disease diagnosis.

Proposal of a grading system for squamous cell carcinoma of the lung - the prognostic importance of tumour budding, single cell invasion, and nuclear diameter.

The prognostic markers of lung squamous cell carcinoma (LSCC) are less investigated. The aim of our study was to evaluate tumour budding (TB), minimal cell nest size, nuclear diameter (ND), and spread through air spaces (STAS) among patients with resected LSCC, semi-quantitatively. Furthermore, we aimed to identify a grading system for the best prognostic stratification of LSCC. Patients who underwent surgical resection at the Department of Surgery, University of Szeged between 2010 and 2016 were included. Follow-up data were collected from medical charts. Morphological characteristics were recorded from histologic revision of slides. Kaplan-Meier analysis, log rank test and Cox proportional-hazards model, ROC curve analysis, and intraclass correlation were utilised. Altogether 220 patients were included. In univariate analysis, higher degree of TB, infiltrative tumour border, larger ND, the presence of single cell invasion (SCI) and STAS were associated with adverse prognosis. Based on our results, we proposed an easily applicable grading scheme focusing on TB, ND, and SCI. In multivariate analysis, the proposed grading system (pOS< 0.001, pRFS< 0.001) and STAS (pOS= 0.008, pRFS< 0.001) were independent prognosticators. Compared to the previously introduced grading systems, ROC curve analysis revealed that the proposed grade had the highest AUC values (AUCOS: 0.83, AUCRFS: 0.78). Each category of the proposed grading system has good (ICC: 0.79-0.88) reproducibility. We validated the prognostic impact of TB, SCI, ND, and STAS in LSCC. We recommend a reproducible grading system combining TB, SCI, and ND for proper prognostic stratification of LSCC patients. Further research is required for validation of this grading scheme.

Cytomorphometric analysis of buccal mucosal cells in iron deficiency anemia: A cross-sectional study.

Worldwide, at any given moment, more individuals have iron deficiency anemia (IDA) than any other health problems. Very few studies have been conducted to evaluate changes in oral mucosa in IDA. The present study is undertaken for cytomorphometric analysis of buccal mucosal cells in IDA. Nuclear diameter (ND), cell diameter (CD), nuclear area (NA), cell area (CA), and nuclear-cytoplasmic ratio (N/C) are measured and compared in buccal mucosal cells of IDA patients and controls. To evaluate cytomorphometric changes in epithelial cells of oral mucosa in IDA and to compare these changes with the controls. A cross-sectional study. Forty cases of IDA and forty cases of control group were selected for the study. IDA cases were diagnosed with decreased Hb level, MCV, MCH, MCHC and confirmed by decreased serum iron and increased total iron-binding capacity levels. Cytomorphometric analysis of buccal mucosa was performed to study CA, CD, NA, ND, and N/C ratio. Statistical analysis was done using t- test in SPSS software version 4. Results of our study showed increased cell area, cell diameter, nuclear area, nuclear diameter, and nuclear-cytoplasmic ratio in anemic group as compared to controls. The study suggests that individuals with IDA regardless of clinically visible oral lesions show cytological changes in oral mucosal epithelium. Knowledge of quantitative alterations in oral epithelial cells of IDA patients is important as these alterations are similar to those seen in precancerous and radiation-induced changes. Cytomorphometry is an efficient tool to understand the extent of cellular changes that occur in oral epithelial cells in IDA.

Effect of fixative solutions in equine ovarian tissue

Histological processing is a classical methodology used for an efficient assessment of ovarian tissue quality. Although it is still controversial which is the most appropriate fixative solution for equine ovarian tissue, the choice of an appropriate fixative is critical to evaluate the improvement of assisted reproductive techniques for fertility preservation. This study aimed to evaluate immediate histological processing, based on an ethanol 70% fixative solution for a short-time exposure (ST-EtOH, 45 min), compared with two classically used fixative solutions [Carnoy (CAR) and paraformaldehyde 4% (PFA)] at different fixation times (6 h, 12 h) on equine ovarian tissue. The end points evaluated were morphology (normal and abnormal) and classes (primordial and developing) of preantral follicles, follicular and stromal cell densities, and follicular and oocyte nuclear diameters. Ovaries (n = 6) from ovariectomized young mares (2–8 years) were fragmented (3 × 3 × 1 mm; 20 fragments/ovary) and fixed in the tested treatments. Overall, a total of 11,661 preantral follicles were evaluated in 1,444 histological slides. High heterogeneity in the number of follicles per individual mare was observed. Equine ovarian fragments fixed in both CAR treatments had a greater (P = 0.014) percentage of morphologically normal primordial and total (primordial and developing classes combined) follicles compared toST-EtOH and both PFA treatments. Although the follicular diameter, regardless of follicular class, was smaller (P = 0.001) in both CAR treatments compared to the other treatments, the nuclear diameter and the nucleus-follicle ratio were greater (P = 0.001) in both CAR treatments compared to the other treatments. A lower (P < 0.001) percentage of morphologically normal developing follicles was observed in ST-EtOH compared to all other treatments. Both CAR treatments had greater (P < 0.001) stromal cell densities than the ST-EtOH and PFA 6h fixatives. In conclusion, Carnoy's solution is the recommended fixative, rather than ST-EtOH and PFA fixatives, for studies requiring more reliable outcomes concerning cellular morphology of equine ovarian tissue. The ST-EtOH fixative proved to be an acceptable alternative to fix equine ovarian tissue when a quick histological evaluation is required for small samples prior to more complicated, time-consuming, and expensive techniques. PFA can be a reasonable fixative for morphological evaluation of equine ovarian tissue, offering the advantage of being suitable for immunohistochemistry techniques since Carnoy's solution is not recommended for this purpose. More studies are required to further understand the effects of different fixative solutions on the morphological ultrastructure and morphometry parameters in different cellular populations of equine ovarian tissue.