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In this study, we evaluated 16 strains of Sarcomyxa edulis from the Changbai Mountains, China through antagonism tests and ISSR molecular marker technique for cytological and molecular biological evaluation. Q-type and R-type cluster analyses were utilised to evaluate and classify agronomic traits according to the test guidelines for distinctness, uniformity, and stability. The tested strains showed antagonistic reactions. The differences in antagonistic reactions among different germplasm resources are helpful for evaluating genetic diversity and provide a basis for better development and utilisation of fungal germplasm resources. A total of 330 bands were amplified by 21 ISSR primers, including 321 polymorphic bands. The percentage of polymorphic bands observed was 97.07%. Using PopGen32 software, the effective number of alleles (Ne), Nei's gene diversity index (H), and Shannon's diversity index (I) were calculated, yielding average values of 1.6638, 0.3796, and 0.5480, respectively. In addition, the genetic similarity coefficient of 16 strains ranged from 0.4306–0.7778. According to the Q-type clustering, the tested strains were classified into two categories based on the Euclidean distance of 8.52. While R-type cluster analysis revealed a significant correlation between the traits. At the Euclidean distance of 6.298, the traits could be classified into three distinct categories. According to the findings, all 16 strains demonstrated high genetic diversity. The ISSR clustering results revealed that strains with shorter genetic distances were more similar in specific traits. However, there were notable differences between the results of comprehensive trait clustering (Q-type clustering) and ISSR clustering. [ABSTRACT FROM AUTHOR]

Background: Human onchocerciasis remains a public health problem in Ghana. Mass drug administration (MDA) with ivermectin (IVM) has reduced disease morbidity and prevalence, but the transmission of onchocerciasis remains ongoing in several endemic foci. We investigated parasite transmission in some endemic communities in Ghana that had received > 18 rounds of annual MDA with IVM and determined the species composition of black fly (Simulium damnosum) vectors in these areas. Methods: Adult female black flies were collected using human landing catches and identified as either forest or savanna species using morpho-taxonomic keys. The adult flies underwent dissection to determine their parity and detect any O. volvulus larvae, followed by the calculation of entomological indices. Simulium damnosum s.l. larvae were collected and preserved in freshly prepared Carnoy's fixative and were later used for cytotaxonomic studies. Results: A total of 9,983 adult flies were caught: 6,569 and 3,414 in the rainy and dry seasons respectively. Black fly biting activities over the study period showed bimodal or trimodal patterns. The highest monthly biting rate (MBR) of 10,578.75 bites/person/month was recorded in July in Beposo, while the highest monthly transmission potential of 100.69 infective bites/person/month was recorded in Asubende in August. Morphological analysis of 2,032 flies showed that 99.8% (2,028) of the flies were savanna species, with only 4 (0.2%) adult flies being of the forest species. Cytogenetic studies on 114 black fly larvae revealed three cytospecies (Simulium damnosum s.s., S. sirbanum and S. sanctipauli) in the study area. Conclusions: The present studies confirmed an ongoing transmission of onchocerciasis in the study communities except Abua-1. It also provides further information on biting behaviors and onchocerciasis transmission indices in the study communities. Further, our data confirmed the savanna species (S. damnosum s.s. and S. sirbanum) of the S.damnosum s.l. to be the major vectors of onchocerciasis in the study areas, with only an occasional influx of forest cytotypes. [ABSTRACT FROM AUTHOR]

Meiosis is a critical process in sexual reproduction, and errors during this cell division can significantly impact fertility. Successful meiosis relies on the coordinated action of numerous genes involved in DNA replication, strand breaks, and subsequent rejoining. DNA topoisomerase enzymes play a vital role by regulating DNA topology, alleviating tension during replication and transcription. To elucidate the specific function of DNA topoisomerase 1α ($AtTOP1 \alpha$ A t T O P 1 α) in male reproductive development of Arabidopsis thaliana, we investigated meiotic cell division in Arabidopsis flower buds. Combining cytological and biochemical techniques, we aimed to reveal the novel contribution of $AtTOP1 \alpha$ A t T O P 1 α to meiosis. Our results demonstrate that the absence of $AtTOP1 \alpha$ A t T O P 1 α leads to aberrant chromatin behavior during meiotic division. Specifically, the top1α1 mutant displayed altered heterochromatin distribution and clustered centromere signals at early meiotic stages. Additionally, this mutant exhibited disruptions in the distribution of 45s rDNA signals and a reduced frequency of chiasma formation during metaphase I, a crucial stage for genetic exchange. Furthermore, the atm-2×top1α1 double mutant displayed even more severe meiotic defects, including incomplete synapsis, DNA fragmentation, and the presence of polyads. These observations collectively suggest that $AtTOP1 \alpha$ A t T O P 1 α plays a critical role in ensuring accurate meiotic progression, promoting homologous chromosome crossover formation, and potentially functioning in a shared DNA repair pathway with ATAXIA TELANGIECTASIA MUTATED (ATM) in Arabidopsis microspore mother cells. [ABSTRACT FROM AUTHOR]

Characteristic volatile organic compounds (VOCs) are anticipated to be used for the identification of lung cancer cells. However, to date, consistent biomarkers of VOCs in lung cancer cells have not been obtained through direct comparison between cancer and healthy groups. In this study, we regulated the glycolysis, a common metabolic process in cancer cells, and employed solid phase microextraction gas chromatography mass spectrometry (SPME–GC–MS) combined with untargeted analysis to identify the characteristic VOCs shared by cancer cells. The VOCs released by three types of lung cancer cells (A549, PC-9, NCI-H460) and one normal lung epithelial cell (BEAS-2B) were detected using SPME–GC–MS, both in their resting state and after treatment with glycolysis inhibitors (2-Deoxy-d-glucose, 2-DG/3-Bromopyruvic acid, 3-BrPA). Untargeted analysis methods were employed to compare the VOC profiles between each type of cancer cell and normal cells before and after glycolysis regulation. Our findings revealed that compared to normal cells, the three types of lung cancer cells exhibited three common differential VOCs in their resting state: ethyl propionate, acetoin, and 3-decen-5-one. Furthermore, under glycolysis control, a single common differential VOC—acetoin was identified. Notably, acetoin levels increased by 2.60–3.29-fold in all three lung cancer cell lines upon the application of glycolysis inhibitors while remaining relatively stable in normal cells. To further elucidate the formation mechanism of acetoin, we investigated its production by blocking glutaminolysis. This interdisciplinary approach combining metabolic biochemistry with MS analysis through interventional synthetic VOCs holds great potential for revolutionizing the identification of lung cancer cells and paving the way for novel cytological examination techniques. [ABSTRACT FROM AUTHOR]

Degradation of shoots, characterized by stunted growth and signs of water deficit, is common in bamboo stands. However, the specific mechanisms underlying water utilization in degrading shoots remain unclear. This study sought to address this gap by harvesting bamboo shoots and culms of Phyllostachys edulis 'Pachyloen', employing cytological and physiological techniques to compare water utilization mechanisms between healthy and degrading shoots, and investigating the water supply to bamboo shoots by the parent bamboo. The water pressure in the degrading shoots was markedly lower compared to that of the healthy shoots, and it declined as the degradation progressed, resulting in reduced water content and the cessation of guttation in the degrading shoots. In conditions of water deficit, the percentage of free water in bamboo shoots decreased while the percentages of bound and semi-bound water increased, with the proportion of semi-bound water reaching as high as 88.13% in the late stages of degradation. The water potential of parent bamboo culms of different ages varied at different times of the day and during different growth stages of bamboo shoots, showing a strong association with the development of bamboo shoots. Conversely, the correlation between changes in the water potential of bamboo shoots and their degradation patterns was found to be comparatively minimal. The weakening of the connection between the bamboo shoots and the parent bamboo culms may play a significant role in the degradation of the bamboo shoots. This is evidenced by a decrease in the fluorescence intensity of the nucleus in bamboo shoots and the degradation of genetic material. This study lays the foundation for future research into the mechanisms of bamboo shoot degradation. [ABSTRACT FROM AUTHOR]

Simple Summary: Various diagnostic methods exist for EC, with traditional biopsy being common but invasive. Endometrial cytology using liquid specimens shows promise for non-invasive detection. Morphological features like fractal dimension, circularity, and solidity in cell clusters could potentially enhance diagnostic accuracy and predict prognosis. Circularity and fractal dimension demonstrated significant associations with EC and AEH, regardless of age and cytology results. An ROC analysis revealed improved diagnostic accuracy when combining fractal dimension with cytology, particularly in menopausal age groups. Moreover, circularity, solidity, and fractal dimension may serve as prognostic indicators for endometrial cancer. This study showed that the morphological characteristics of cell clusters in liquid-based cytology may improve endometrial cancer diagnosis and predict prognosis. Endometrial cancer (EC) in women is increasing globally, necessitating improved diagnostic methods and prognosis prediction. While endometrial histology is the conventional approach, liquid-based endometrial cytology may benefit from novel analytical techniques for cell clusters. A clinical study was conducted at the University of Fukui Hospital from 2012 to 2018, involving 210 patients with endometrial cytology. The liquid-based cytology images were analyzed using cell cluster analysis with Image J software. Logistic regression, ROC analysis, and survival analysis were employed to assess the diagnostic accuracy and prognosis between cell cluster analysis and EC/atypical endometrial hyperplasia (AEH). Circularity and fractal dimension demonstrated significant associations with EC and AEH, regardless of age and cytology results. The ROC analysis revealed improved diagnostic accuracy when combining fractal dimension with cytology, particularly in menopausal age groups. Lower circularity and solidity were independently associated with poor overall survival, while higher fractal dimension values correlated with poorer overall survival in Grades 2 and 3 endometrial cancers. The combination of circularity and fractal dimension with cytology improved diagnostic accuracy for both EC and AEH. Moreover, circularity, solidity, and fractal dimension may serve as prognostic indicators for endometrial cancer, contributing to the development of more refined screening and diagnostic strategies. [ABSTRACT FROM AUTHOR]

Objective: The Visiopharm artificial intelligence (AI) algorithm for oestrogen receptor (ER) immunohistochemistry (IHC) in whole slide images (WSIs) has been successfully validated in surgical pathology. This study aimed to assess its efficacy in cytology specimens. Methods: The study cohort comprised 105 consecutive cytology specimens with metastatic breast carcinoma. ER IHC WSIs were seamlessly integrated into the Visiopharm platform from the Image Management System (IMS) during our routine digital workflow, and an AI algorithm was employed for analysis. ER AI scores were compared with pathologists' manual consensus scores. Optimization steps were implemented and evaluated to reduce discordance. Results: The overall concordance between pathologists' scores and AI scores was excellent (99/105, 94.3%). Six cases exhibited discordant results, including two false‐negative (FN) cases due to abundant histiocytes incorrectly counted as negatively stained tumour cells by AI, two FN cases owing to weak staining, and two false‐positive (FP) cases where pigmented macrophages were erroneously counted as positively stained tumour cells by AI. The Pearson correlation coefficient of ER‐positive percentages between pathologists' and AI scores was 0.8483. Optimization steps, such as lowering the cut‐off threshold and additional training using higher input magnification, significantly improved accuracy. Conclusions: The automated ER AI algorithm demonstrated excellent concordance with pathologists' assessments and accurately differentiated ER‐positive from ER‐negative metastatic breast carcinoma cytology cases. However, precision in identifying tumour cells in cytology specimens requires further enhancement. [ABSTRACT FROM AUTHOR]

Thyroid cancer diagnosis primarily relies on imaging techniques and cytological analyses. In cases where the diagnosis is uncertain, the quantification of molecular markers has been incorporated after cytological examination. This approach helps physicians to make surgical decisions, estimate cancer aggressiveness, and monitor the response to treatments. Despite the availability of commercial molecular tests, their widespread use has been hindered in our experience due to cost constraints and variability between them. Thus, numerous groups are currently evaluating new molecular markers that ultimately will lead to improved diagnostic certainty, as well as better classification of prognosis and recurrence. In this review, we start reviewing the current preoperative testing methodologies, followed by a comprehensive review of emerging molecular markers. We focus on micro RNAs, long non-coding RNAs, and mitochondrial (mt) signatures, including mtDNA genes and circulating cell-free mtDNA. We envision that a robust set of molecular markers will complement the national and international clinical guides for proper assessment of the disease. [ABSTRACT FROM AUTHOR]

Background & Aims: Endoscopic retrograde cholangiopancreatography (ERCP) with brush cytology is an important tool in the diagnosis of hepatobiliary malignancies. However, reported sensitivity of brush cytology is suboptimal and differs markedly per study. The aim of this study is to analyze the optimal technique of endobiliary brushing during ERCP. Methods: A systematic review and meta-analysis according was performed using Pubmed, Embase and Cochrane library, and reported reported according to the PRISMA guidelines. The intervention reported should involve ERCP, performed by the endoscopist with a comparison of different brushing techniques. The primary outcome was sensitivity for malignancy. Studies published up to December 2022 were included. Percutaneous techniques and cytological or laboratory techniques for processing of material were excluded. Bias was assessed using the Quadas-2 tool. Pooled sensitivity rates and Forest plots were analyzed for the primary outcome. Results: A total of 16 studies were included. Three studies reported on brushing before or after dilation of a biliary stricture. No improvement in sensitivity was found. Five studies reported on alternative brush designs. This did not lead to improved sensitivity. Seven studies reported on the aspiration and analysis of bile fluid, which resulted in a 16% increase in sensitivity (95% CI 4-29%). One study reported an increased in the number of brush passes to the stricture, providing an increase in sensitivity of 20%. Substantial heterogeneity between studies was found, both methodological and statistical. Conclusions: Increasing the number of brush-passes and sending bile fluid for cytology increases the sensitivity of biliary brushings during ERCP. Dilation before brushing or alternative brush designs did not increase sensitivity. [ABSTRACT FROM AUTHOR]

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Papillary thyroid cancer (PTC) is a common endocrine malignancy, and its incidence is reported to be constantly increasing. BRAF mutation is detected in approximately 44% of PTCs, and the most common BRAF mutation is thymine (T) to adenine (A) missense mutation in nucleotide 1796 (T1796A, V600E). Although BRAFV600E represents 95% of all BRAF mutations, uncommon BRAF mutations have been reported in thyroid carcinomas and represent an alternative mechanism of BRAF activation with unclear clinical significance. We report a novel non-V600E mutation (c.1799_1812delinsAT, p.V600_W604delinsD), identified preoperatively with next-generation sequencing (NGS) on the material obtained with fine-needle aspiration cytology (FNAC) performed on a thyroid nodule cytologically suspicious for malignancy in a 35-year-old male patient. The presence of this new variant of BRAF mutation was subsequently confirmed in the postoperative phase by direct Sanger sequencing. In conclusion, we report a new non-V600E variant previously undetected in papillary thyroid cancer. In addition, this case report shows that the NGS technique on cytological tissue allows to detect the presence of rare mutations, thus increasing the diagnostic specificity of molecular analysis. [ABSTRACT FROM AUTHOR]

Introduction: Cervical cancer is the third leading cause of death from malignancies worldwide, affecting mainly low- and middle-income countries. By 2020 an incidence of 604,000 new cases was estimated. Objective: To characterize the main hospital indicators of the Cervical Cancer Early Diagnosis Program. Methods: An observational, descriptive and cross-sectional study was carried out to characterize the main hospital indicators of the Program in the Neck Pathology Service of the General Hospital Dr. Bruno Zayas Alfonso of Santiago de Cuba, from January 2020 to December 2022. The study population consisted of 443 women. The data collected were analyzed using descriptive statistic techniques, expressed in frequency and percentages. Results: Of the women studied, 60.9 %had cervical intraepithelial lesion of high degree of malignancy, with 32.6 % positive for cervical cancer. The overall percentage of useless cytological tests was 2.07 and no cells from the processing zone was 4.01, both quality indicators. There was a high significance in terms of the overall coverage rate of women at risk (K=0.615), showing that the results of the cytology largely reflect the diagnosis of histology, with a good agreement. Conclusions: Cervical-vaginal cytology remains the most valuable diagnostic method for detecting cervical intraepithelial neoplasm and early-stage carcinoma in large populations [ABSTRACT FROM AUTHOR]

Neurodegeneration arising from aging, injury, or diseases has devastating health consequences. Whereas neuronal survival and axon degeneration have been studied extensively, much less is known about how neurodegeneration affects dendrites, in part due to the limited assay systems available. To develop an assay for dendrite degeneration and repair, we used photo-switchable caspase-3 (caspase-Light-Oxygen-Voltage-sensing [caspase-LOV]) in peripheral class 4 dendrite arborization (c4da) neurons to induce graded neurodegeneration by adjusting illumination duration during development and adulthood in Drosophila melanogaster. We found that both developing and mature c4da neurons were able to survive while sustaining mild neurodegeneration induced by moderate caspase-LOV activation. Further, we observed active dendrite addition and dendrite regeneration in developing and mature c4da neurons, respectively. Using this assay, we found that the mouse Wallerian degeneration slow (WldS) protein can protect c4da neurons from caspase-LOV-induced dendrite degeneration and cell death. Furthermore, our data show that WldS can reduce dendrite elimination without affecting dendrite addition. In summary, we successfully established a photo-switchable assay system in both developing and mature neurons and used WldS as a test case to study the mechanisms underlying dendrite regeneration and repair.

Background and Aim: A combined microbial and cytological examination of uterine samples is the main diagnostic method for endometritis in mares. This study aimed to describe a procedure for using the same uterine cytobrush (CB) for both bacteriological and cytological evaluation. Material and Methods: The procedure consists of rolling the CB onto a sterilized glass slide immediately after collection and before the transfer into a sterile saline solution. In Experiment 1, a comparison between bacteriological results of the cotton swab (CS) and CB or pellet was made in 10 mares; in Experiment 2, bacteriological and cytological results were compared between different processing methods of CB in 28 mares; in other 6 mares, a CB was processed for cytology only, to investigate the reasons for the low cellularity of the pellet. Results: The agreement between culture results from the CB and CS was evaluated, and a comparison between the cytological data obtained by different processing methods of CB was performed. The perfect agreement between the CB and CS microbiological results was found. The described procedure enables useful diagnostic smears for cytology. Moreover, the seeding of both the tip of CB and the saline solution used for the transport produced accurate bacteriological results. Conclusion: The protocol described in this study for the use of CB for both cytological and bacteriological analysis could be used for the diagnosis of endometritis. To maximize diagnostic sample quality, cytology slides must be prepared with meticulous care in the field to preserve cellular integrity and minimize artifacts. [ABSTRACT FROM AUTHOR]

Objective: The proposed study was planned to screen Human Papilloma Virus (HPV) status in potentially malignant oral disorders (PMOD) and correlated HPV positivity with cytological changes in oral smears. Methods: This descriptive cross-sectional study was conducted at University of Health Sciences Lahore, Pakistan from April 2020 to April 2021. Oral smears from N=162 patients with PMODs were taken by the Cytobrush and Manual Liquid Based Cytology was performed followed by p16 antibody detection on immunohistochemistry and HPV-DNA detection by conventional polymerase chain reaction (PCR). The cytological changes were categorized according to the updated Bethesda Classification system 2014. SPSS was used to analyze data and p-Value of <0.05 was considered as statistically significant. Results: Out of total N = 162 patients, the most prevalent lesion [39% (n=63)] was lichen planus. Fifty six percent (n=90) of the patients were habitual chewers and 43% (n=70) were smokers. Pap staining of oral smears revealed atypical squamous cells of undetermined significance (ASCUS) in 45% (n=69) cases and in 2 % (n=4) of the samples diagnosis of atypical squamous cells-cannot exclude high-grade squamous intraepithelial lesion (ASC-H) was made. A total of 37% cases showed HPV positivity by polymerase chain reaction (PCR) while positive p16 expression was observed in 24% of the cases. ASC-H and ASCUS category showed significant association with HPV positivity (p=0.003). Conclusion: Early detection of PMODs by adopting minimally invasive cytological techniques and screening for HPV infection in local population is pivotal to reduce the morbidity and mortality associated with the advanced disease and carcinoma. [ABSTRACT FROM AUTHOR]

Introduction: Specific diagnosis of endometrial carcinomas on cervical cytology is difficult with few useful cytomorphological clues reported. This study reviews a cohort of cervical cytology to investigate the presence of keratinization in atypical glandular cells (AGC), an undescribed cytomorphological clue for identifying endometrial endometrioid carcinomas on cervical cytology. Methods: Cervical cytology slides from patients with a histologic diagnosis of endometrial endometrioid carcinoma were reviewed for the presence of keratinization associated with AGCs. Corresponding histology slides were reviewed for tumour grading and degree of squamous differentiation. Results: In total, 42 cases of cervical cytology specimens from 41 patients were retrieved, including 7 (16.7%) with keratinization associated with AGCs seen and 35 (83.3%) without. Comparison of histologic grading did not demonstrate an association with the presence of keratinization on cytology (p = 0.565). Corresponding histology slides were available for 37 cases. Cytologic and histologic keratinization were associated statistically (p = 0.002). Frank keratinization was seen on histologic slides of five cases, with four also showing cytologic keratinization. Area of squamous differentiation, including squamous morule formation, did not correlate with keratinization on cytologic preparation (p = 0.185). Conclusion: Histologic and cytologic keratinization are observed in endometrioid endometrial carcinomas. Such is reflected in cervical cytology by the presence of orangeophilic, rigid and acellular fragments within or associated with AGC clusters. Keratinization, when identified with AGCs, should be regarded as a cytologic clue suggestive of an endometroid carcinoma of endometrial origin. [ABSTRACT FROM AUTHOR]

A five‐year‐old male English Bulldog was presented with a firm, well‐circumscribed, 1 cm in diameter cutaneous mass on the left flank. Fine‐needle aspiration (FNA) biopsy samples were collected for cytologic analysis. Cytology revealed a highly cellular sample consisting of spindle cells, numerous bundles of thick, glassy eosinophilic material (hyalinized collagen), and inflammatory cells. Spindle cells showed moderate anisocytosis and anisokaryosis, had oval nuclei with coarsely stippled chromatin, 1–3 prominent round nucleoli, and moderate amounts of wispy cytoplasm. Cells were occasionally associated with an eosinophilic extracellular matrix. Binucleated and trinucleated spindle cells were often noted. Low numbers of macrophages, small lymphocytes, and individual well‐granulated mast cells were also present. The lesion was excised and submitted for histopathologic examination, revealing a well‐delineated, nonencapsulated mass composed of hyalinized collagen fibers separated by spindle‐shaped mesenchymal cells in the deep dermis and subcutis. Mild anisocytosis and anisokaryosis and less than one mitosis per 10 × high power fields were present. Excision of the mass was complete. The findings were consistent with a keloidal fibroma, a rare benign variant of fibroma. Neoplastic cells showed positive immunoreactivity for vimentin, and a small‐to‐moderate number of tumor cells showed positive immunoreactivity for α‐smooth muscle actin. This is the first cytologic description of a keloidal fibroma correlated with histopathologic findings and immunolabeling. In cases where keloidal neoplasia is suspected, and since moderate cellular atypia can be present on cytologic examination even in cases of keloidal fibroma, histopathologic examination is necessary to differentiate between keloidal fibroma and keloidal fibrosarcoma. [ABSTRACT FROM AUTHOR]

Background: Collecting cytology samples and making simple diagnoses are skills taught in veterinary universities, mostly in a passive way. Simulators enhance practical skills learning, increasing student engagement through immersive activities. These strategies have not been carefully assessed in veterinary cytology. Objectives: This study aimed to describe a simulator developed for training cytology sample collection methods and assess the utility of an immersive simulation strategy to learn and practice the collection of cytology samples. Methods: A flipped classroom with a station design was followed. Students first watched video tutorials on sampling collection, listened to brief cases' clinical histories, and moved to immersive simulator stations. There, they practiced imprints, swabs, and fine‐needle aspiration (FNA). Microscopic observation of the material was available through augmented reality tools. Students were evaluated by Objective Structured Clinical Examination (OSCE) tools on their ability to perform FNA on stuffed animal models. Students answered two questionnaires addressing their learning outcomes. Results: Second‐ and third‐year students from two centers (n = 129) practiced cytologic collection methods in simulators which significantly changed their willingness to perform FNA in live animals after the class activities. OSCE pass rates over 90% were obtained for most steps of FNA, and students rated the activity as essential/very relevant for learning. Conclusions: Immersive simulation strategies were effective at increasing student comfort with cytologic sampling techniques. This approach should be included in the veterinary curriculum as it can increase the quality of cytology samples and could potentially improve the cytologic diagnosis of a submitted sample. [ABSTRACT FROM AUTHOR]

Background: Distinguishing primary and secondary pulmonary neoplasms can be challenging via cytology, and a rapid, inexpensive diagnostic tool to differentiate these neoplasms is unavailable. Alkaline phosphatase cytochemistry (ALP‐CC) has been used to identify primary pulmonary carcinomas in human patients, and we hypothesized it could be applied to canine lung aspirates. Objective: We aimed to characterize ALP‐CC expression in fine‐needle aspirate (FNA) samples of canine pulmonary neoplastic and non‐neoplastic tumors. Methods: A retrospective case search was conducted to identify cases with contemporaneous cytology and histopathology reports from pulmonary lesions, including neoplastic and non‐neoplastic etiologies. Slides prepared from pulmonary aspirates were stained for ALP‐CC activity, and the percentage of ALP‐CC‐positive primary pulmonary epithelial tumors was determined. To characterize the ALP‐CC expression in non‐neoplastic cellular constituents of pulmonary FNA samples, mesothelial cells were also evaluated. Results: Forty‐eight canine cases met the inclusion criteria. ALP‐CC–positive cells were seen in both neoplastic and non‐neoplastic lesions. In non‐neoplastic lesions, pulmonary epithelial cells were ALP‐CC positive. Eighty‐nine percent of primary pulmonary epithelial neoplasms were ALP‐CC positive, and no ALP‐CC positivity was noted in mesothelial cells. ALP‐CC–positive neoplastic cells were seen in a metastatic amelanotic melanoma. Conclusions: Primary pulmonary epithelial neoplasms are frequently ALP‐CC positive, but such positivity is not restricted to this tumor type. Non‐neoplastic pulmonary epithelial cells can be ALP‐CC positive, whereas mesothelial cells are negative. [ABSTRACT FROM AUTHOR]

The World Health Organization (WHO) has recently published a new lung cytopathology reporting system, which simplifies the diagnostic categories into a five-tier system without subcategorization. The system encourages the use of ancillary diagnostic tests for better categorization of specimens and further management. This correspondence provides a brief outline of the lung reporting system, including standardized nomenclature and the use of terminologies harmonizing with the WHO Blue Book. The system includes categories such as non-diagnostic, benign, atypical, suspicious for malignancy, and malignant, with corresponding risk-based management strategies for clinicians. Ancillary techniques like immunocytochemistry can assist in upgrading the category from suspicious for malignancy to malignancy. The system aims to provide universal simplified criteria for categorizing lung cytology samples and their management options. [Extracted from the article]

BackgroundMen who have sex with men (MSM) living with human immunodeficiency virus (HIV) are at increased risk of anal cancer. Anal cytology can be used to screen for dysplasia, with high-resolution anoscopy (HRA) required for diagnostic confirmation. We describe the impact lack of HRA had on management of abnormal screening results in Bogotá, Colombia.Material and methodsThis retrospective cohort study includes MSM with HIV who underwent anal cytology screening between January 2019February 2020, with colorectal surgery (CRS) follow-up through July 2020. Cytology results included atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), and high-grade squamous intraepithelial lesion (HSIL). Categorical and continuous variables were compared via Fisher's exact test and Wilcoxon rank-sum, respectively.ResultsOf 211 MSM screened, 68 had abnormal cytology: ASC-US (n = 23), LSIL (n = 41), HSIL (n = 4). Sixty (88.2%) were referred to CRS, and 51 (75.0%) attended ≥ 1 appointment. At initial assessment, 17 were referred for anal exam under anesthesia (EUA) for tissue resection, and 21 for rectosigmoidoscopy. Having perianal condyloma was associated with recommendation for EUA (P < 0.001), while cytology grade of dysplasia was not (P = 0.308). Eleven (16.2%) underwent EUA for condyloma resection.ConclusionsFew studies have described anal cancer screening in settings without HRA. We found lack of HRA limited management of abnormal cytology in Colombia. Those with condyloma underwent resection, but HRA remains necessary to localize and treat microscopic disease. Next steps include implementation of HRA in order to further develop the anal cancer screening program for MSM with HIV in Bogotá.

ObjectiveTo assess correlations between clinical and cytological features of feline eosinophilic keratoconjunctivitis at the time of cytological diagnosis.Animals studiedFifteen client-owned, domestic breed cats (18 eyes) examined between 2007 and 2019.ProceduresAn electronic search and medical record review of cats diagnosed with feline eosinophilic keratitis or keratoconjunctivitis (FEK) based on clinical examination findings and eosinophils detected on corneal cytology were conducted. Clinical severity was graded using a modified version of a previously validated semiquantitative preclinical ocular toxicology scoring (SPOTS) system. Clinical grades were assigned following review of clinical images and medical record descriptions, and cytological grades were assigned following review of archived corneal cytology slides. Correlations were analyzed for significance using Spearman's rank correlation coefficient.ResultsHigher total corneal scores correlated with higher total conjunctival scores, but not with total fluorescein scores. Small lymphocyte scores correlated negatively with scores for collagen degeneration or mineralization. Globule leukocytes, a unique cell type not previously described in ocular cytology, were identified in 4 of 18 cytological samples. Higher globule leukocyte scores were correlated with higher scores for mast cells or plasma cells. Specimens with lower eosinophil scores had higher globule leukocyte scores.ConclusionsLarge variability was detected in the cytological characteristics and clinical features of FEK-affected cats. This is the first report of globule leukocytes being identified in ocular cytology from any species. The role of globule leukocytes in the etiopathogenesis and progression of FEK remains unknown and warrants further investigation.

Objective: A case of intestinal spirochaetosis diagnosed in anal cytology is presented. Methods: An anal liquid‐based cytology was performed and stained with Papanicolau and Warthin‐Starry stains. Results: An apical filamentous "fringe" was identified in columnar cells groups. Conclusion: Anal cytology is a potential tool for intestinal spirochaetosis diagnosis. [ABSTRACT FROM AUTHOR]

Background: To use different cytological staining techniques to examine the cytomorphologic aspects of thyroid lesions. Materials and Methods: 100 individuals with adequate aspirates participated in an experimental study carried out at the cytopathology outpatient department. The assignment was finished when the Institutional Ethical Committee granted the necessary authorization. Results: Histopathological correlation was obtained for 25 of the 100 cases in this investigation. For seven cases of Hashimoto's thyroiditis, one case of lymphocytic thyroiditis, six cases of nodular colloid goitre, eight cases of nodular colloid goitre with cystic degeneration, seven cases of follicular neoplasm, and six cases of thyroid papillary carcinoma, histopathological linkage was available. Histopathological studies of those 25 instances showed a correlation with the cytological diagnosis. Three instances that were first diagnosed as follicular neoplasms based on cytology were later identified as follicular carcinomas. Conclusion: A combination of several stains and different fixation procedures will boost the diagnostic accuracy of thyroid lesions. Thus, in order to decrease false positive and false negative results in fine aspiration cytology of thyroid lesions, a range of cytological staining methods can be applied, given appropriate material availability. [ABSTRACT FROM AUTHOR]

Purpose: Thyroid nodules classified as indeterminate in previous fine-needle aspiration cytology often necessitate additional evaluation to determine their histology, while shear wave elastography (SWE) offers an alternative option in this regard. The objective of this study was to assess the diagnostic effectiveness of SWE in evaluating indeterminate nodules. Methods: The PubMed, EMBASE, and Web of Science databases were searched from 1st January 1970 to 1st March 2023. The studies were reviewed and the data was extracted by two separate reviewers. A Bayesian bivariate model was utilized to quantitatively synthesize the diagnostic accuracy and yield of the studies in R. Results: A total of seven studies, involving indeterminate thyroid nodules undergoing SWE were included, and the overall malignancy rate was 34.1% (307/900). The summarized estimates of sensitivity and specificity were 0.792 (95% credible interval [CI], 0.727–0.850) and 0.845 (95% CI, 0.797–0.887), respectively. The summarized estimate for the diagnostic odds ratio (DOR) was 17.8 (95% CI, 14.0–22.6). Summarized receiver operating characteristic (SROC) plots indicated a trade-off between sensitivity and specificity, and the estimate of AUC was 0.866 (95% CI, 0.834–0.895). The summary estimates for positive and negative likelihood ratios were 4.67 (95% CI, 3.98–5.85) and 0.26 (95% CI, 0.23–0.28), respectively. Conclusions: The overall accuracy of SWE remains satisfactory in indeterminate thyroid nodules. However, it should be noted that the available data are still extremely limited, and more studies or guidelines are required to provide further insights.

Objectives and Background: To use a variety of cytological staining techniques to examine the cytomorphologic aspects of thyroid lesions. Single thyroid nodules and multinodular goiters are both equally common outside of cancer hotspots, and both have about the same risk of becoming malignant. Methods: This is a prospective experimental study carried out at Department of Pathology, Teritary Care Hospital, Bangalore, India, over the period of February 2022 to February 2023. 70 cases with adequate aspirates were considered for this study. Results: The cytomorphological features of individual thyroid lesions are analyzed in this study using H&E, Pap and Wright Giemsa stains. Many cytology facilities use these three stains routinely. 22 cases of Hashimoto's thyroiditis, 4 cases of lymphocytic thyroiditis, 25 cases of Nodular colloid goitre, 9 cases of Nodular colloid goitre with cystic change, 5 cases of follicular neoplasm, 1 case of suspicious of papillary carcinoma of thyroid, 4 cases of papillary carcinoma of thyroid patients were surveyed for this study. Pap stained smears make follicular cells, which have powdered chromatin and tiny nucleoli, easy to see. Wright Giemsa stained thyroid tissue smears exhibited follicular cells with enlarged nuclei, uneven nuclear outlines, small granular chromatin and dilated cytoplasms. Cell nuclei had cytoplasmic inclusions. Conclusion: Thyroid lesion diagnosis is greatly aided, in our opinion, by the use of a wide range of fixation and staining methods. Hence, if enough tissue is available, fine aspiration cytology of thyroid lesions can use a variety of cytological staining techniques to limit false positive and false negative results. [ABSTRACT FROM AUTHOR]

The emergence of antibiotic resistance in bacteria has led to the investigation of alternative treatments, such as phage therapy. In this study, we examined the interactions between the nucleus-forming jumbo phage ФKZ and antibiotic treatment against Pseudomonas aeruginosa. Using the fluorescence microscopy technique of bacterial cytological profiling, we identified mechanism-of-action-specific interactions between antibiotics that target different biosynthetic pathways and ФKZ infection. We found that certain classes of antibiotics strongly inhibited phage replication, while others had no effect or only mildly affected progression through the lytic cycle. Antibiotics that caused an increase in host cell length, such as the cell wall active antibiotic ceftazidime, prevented proper centering of the ФKZ nucleus via the PhuZ spindle at midcell, leading us to hypothesize that the kinetic parameters of the PhuZ spindle evolved to match the average length of the host cell. To test this, we developed a computational model explaining how the dynamic properties of the PhuZ spindle contribute to phage nucleus centering and why some antibiotics affect nucleus positioning while others do not. These findings provide an understanding of the molecular mechanisms underlying the interactions between antibiotics and jumbo phage replication. [ABSTRACT FROM AUTHOR]

Aims: Thyroid nodules are one of the most frequent medical issues in endocrinology in our country and around the world. The appropriate evaluation of the nodule is critical in the management of patients. Fine needle aspiration cytology (FNAC) is one of the most accurate tools for evaluating these nodules. Conventional and liquid-based (LB) methods are available for thyroid FNAC. In this paper, we aim to determine the best cytological method for the evaluation of thyroid fine needle aspiration materials. Settings and Design: Fine needle aspiration (FNA) materials obtained and prepared by conventional and LB methods from 310 patients were evaluated. These slides were compared in terms of 12 cytological parameters in a three-tiered system, and the kappa coefficient was calculated. Methods and Material: Two slides were prepared from FNA samples, and the rest of the material was left in protective solution for LB cytology. Surepath was used as a LB technique. Conventional slides (CS) were left to dry in the air and stained with May Grunwald Giemsa. Slides prepared by two different techniques were compared in 12 cytomorphological parameters, and Kappa coefficients and correlations were calculated. Statistical Analysis Used: Kappa statistics. Results: In comparison of LB and CS, the highest kappa coefficient was in chromatin texture (k:0.738) and inflammatory cells (k: 0.482). On other parameters, fair or poor agreement was observed. Conclusions: Although there are some superiorities to LB cytology, co-application of these two methods is more favorable in thyroid fine needle aspiration. [ABSTRACT FROM AUTHOR]

Objective: Artificial intelligence (AI)–based cytopathology studies conducted using deep learning have enabled cell detection and classification. Liquid‐based cytology (LBC) has facilitated the standardisation of specimen preparation; however, cytomorphology varies according to the LBC processing technique used. In this study, we elucidated the relationship between two LBC techniques and cell detection and classification using a deep learning model. Methods: Cytological specimens were prepared using the ThinPrep and SurePath methods. The accuracy of cell detection and cell classification was examined using the one‐ and five‐cell models, which were trained with one and five cell types, respectively. Results: When the same LBC processing techniques were used for the training and detection preparations, the cell detection and classification rates were high. The model trained on ThinPrep preparations was more accurate than that trained on SurePath. When the preparation types used for training and detection were different, the accuracy of cell detection and classification was significantly reduced (P < 0.01). The model trained on both ThinPrep and SurePath preparations exhibited slightly reduced cell detection and classification rates but was highly accurate. Conclusions: For the two LBC processing techniques, cytomorphology varied according to cell type; this difference affects the accuracy of cell detection and classification by deep learning. Therefore, for highly accurate cell detection and classification using AI, the same processing technique must be used for both training and detection. Our assessment also suggests that a deep learning model should be constructed using specimens prepared via a variety of processing techniques to construct a globally applicable AI model. [ABSTRACT FROM AUTHOR]

Multiplex protein analysis and ensemble machine learning methods of fine needle aspirates fro... I Pontus Röbeck, Bo Franzén, Rafaele Cantera-Ahlman, Anca Dragomir, Gert Auer, Håkan Jorulf, Sven P. Jacobsson, Kristina Viktorsson, Rolf Lewensohn, Michael Häggman, Sam Ladjevardi i DOI: 10.1111/cyt.13226 | Version of Record Online: 20 March 2023 This pilot study shows that multiplex profiling of potential diagnostic biomarkers and immunotherapy targets is feasible on tumour materials obtained by fine needle aspiration sampling of prostate cancer. Atypical spindle cell/pleomorphic lipomatous tumour (ASPLT): A report of three FNA cases and... I Paul E. Wakely Jr i DOI: 10.1111/cyt.13227 | Version of Record Online: 17 March 2023 Atypical spindle cell/pleomorphic lipomatous tumor (ASPLT) is a newly classified adipocytic neoplasm whose cytopathology has not been previously characterized. Cytological findings in a case of endocrine mucin-producing sweat gland carcinoma with metast... I Min-Hsiang Chang, Chien-Hui Wu i DOI: 10.1111/cyt.13239 | Version of Record Online: 26 April 2023 Endocrine mucin-producing sweat gland carcinoma (EMPSGC) was previously considered an indolent skin tumour without metastasis. [Extracted from the article]

Background: In diseases such as interstitial lung diseases (ILDs), patient diagnosis relies on diagnostic analysis of bronchoalveolar lavage fluid (BALF) and biopsies. Immunological BALF analysis includes differentiation of leukocytes by standard cytological techniques that are labor-intensive and time-consuming. Studies have shown promising leukocyte identification performance on blood fractions, using third harmonic generation (THG) and multiphoton excited autofluorescence (MPEF) microscopy. Objective: To extend leukocyte differentiation to BALF samples using THG/MPEF microscopy, and to show the potential of a trained deep learning algorithm for automated leukocyte identification and quantification. Methods: Leukocytes from blood obtained from three healthy individuals and one asthma patient, and BALF samples from six ILD patients were isolated and imaged using label-free microscopy. The cytological characteristics of leukocytes, including neutrophils, eosinophils, lymphocytes, and macrophages, in terms of cellular and nuclear morphology, and THG and MPEF signal intensity, were determined. A deep learning model was trained on 2D images and used to estimate the leukocyte ratios at the image-level using the differential cell counts obtained using standard cytological techniques as reference. Results: Different leukocyte populations were identified in BALF samples using label-free microscopy, showing distinctive cytological characteristics. Based on the THG/MPEF images, the deep learning network has learned to identify individual cells and was able to provide a reasonable estimate of the leukocyte percentage, reaching >90% accuracy on BALF samples in the hold-out testing set. Conclusions: Label-free THG/MPEF microscopy in combination with deep learning is a promising technique for instant differentiation and quantification of leukocytes. Immediate feedback on leukocyte ratios has potential to speed-up the diagnostic process and to reduce costs, workload and inter-observer variations. [ABSTRACT FROM AUTHOR]

Aims: Thyroid nodules are one of the most frequent medical issues in endocrinology in our country and around the world. The appropriate evaluation of the nodule is critical in the management of patients. Fine needle aspiration cytology (FNAC) is one of the most accurate tools for evaluating these nodules. Conventional and liquid-based (LB) methods are available for thyroid FNAC. In this paper, we aim to determine the best cytological method for the evaluation of thyroid fine needle aspiration materials. Settings and Design: Fine needle aspiration (FNA) materials obtained and prepared by conventional and LB methods from 310 patients were evaluated. These slides were compared in terms of 12 cytological parameters in a three-tiered system, and the kappa coefficient was calculated. Methods and Material: Two slides were prepared from FNA samples, and the rest of the material was left in protective solution for LB cytology. Surepath was used as a LB technique. Conventional slides (CS) were left to dry in the air and stained with May Grunwald Giemsa. Slides prepared by two different techniques were compared in 12 cytomorphological parameters, and Kappa coefficients and correlations were calculated. Statistical Analysis Used: Kappa statistics. Results: In comparison of LB and CS, the highest kappa coefficient was in chromatin texture (κ:0.738) and inflammatory cells (κ: 0.482). On other parameters, fair or poor agreement was observed. Conclusions: Although there are some superiorities to LB cytology, co-application of these two methods is more favorable in thyroid fine needle aspiration.

The idea that liquid-liquid phase separation (LLPS) may be a general mechanism by which molecules in the complex cellular milieu may self-organize has generated much excitement and fervor in the cell biology community. While this concept is not new, its rise to preeminence has resulted in renewed interest in the mechanisms that shape and drive diverse cellular self-assembly processes from gene expression to cell division to stress responses. In vitro biochemical data have been instrumental in deriving some of the fundamental principles and molecular grammar by which biological molecules may phase separate, and the molecular basis of these interactions. Definitive evidence is lacking as to whether the same principles apply in the physiological environment inside living cells. In this Perspective, we analyze the evidence supporting phase separation in vivo across multiple cellular processes. We find that the evidence for in vivo LLPS is often phenomenological and inadequate to discriminate between phase separation and other possible mechanisms. Moreover, the causal relationship and functional consequences of LLPS in vivo are even more elusive. We underscore the importance of performing quantitative measurements on proteins in their endogenous state and physiological abundance, as well as make recommendations for experiments that may yield more conclusive results.

BackgroundLittle is known about how bacterial endosymbionts colonize host tissues. Because many insect endosymbionts are maternally transmitted, egg colonization is critical for endosymbiont success. Wolbachia bacteria, carried by approximately half of all insect species, provide an excellent model for characterizing endosymbiont infection dynamics. To date, technical limitations have precluded stepwise analysis of germline colonization by Wolbachia. It is not clear to what extent titer-altering effects are primarily mediated by growth rates of Wolbachia within cell lineages or migration of Wolbachia between cells.ResultsThe objective of this work is to inform mechanisms of germline colonization through use of optimized methodology. The approaches are framed in terms of nutritional impacts on Wolbachia. Yeast-rich diets in particular have been shown to suppress Wolbachia titer in the Drosophila melanogaster germline. To determine the extent of Wolbachia sensitivity to diet, we optimized 3-dimensional, multi-stage quantification of Wolbachia titer in maternal germline cells. Technical and statistical validation confirmed the identity of Wolbachia in vivo, the reproducibility of Wolbachia quantification and the statistical power to detect these effects. The data from adult feeding experiments demonstrated that germline Wolbachia titer is distinctly sensitive to yeast-rich host diets in late oogenesis. To investigate the physiological basis for these nutritional impacts, we optimized methodology for absolute Wolbachia quantification by real-time qPCR. We found that yeast-rich diets exerted no significant effect on bodywide Wolbachia titer, although ovarian titers were significantly reduced. This suggests that host diets affects Wolbachia distribution between the soma and late stage germline cells. Notably, relative qPCR methods distorted apparent wsp abundance, due to altered host DNA copy number in yeast-rich conditions. This highlights the importance of absolute quantification data for testing mechanistic hypotheses.ConclusionsWe demonstrate that absolute quantification of Wolbachia, using well-controlled cytological and qPCR-based methods, creates new opportunities to determine how bacterial abundance within the germline relates to bacterial distribution within the body. This methodology can be applied to further test germline infection dynamics in response to chemical treatments, genetic conditions, new host/endosymbiont combinations, or potentially adapted to analyze other cell and tissue types.

Introduction: Despite the fact that Fine Needle Aspiration Cytology (FNAC) has been widely utilised in the preoperative diagnosis of breast lumps, the Conventional Smears (CS) have drawbacks, including difficulty in understanding the pattern or architecture of the lesion, determining invasiveness, Immunohistochemistry (IHC), false positives, and false negatives. Cytologists advise using Cell Blocks (CB) to increase the diagnostic precision of FNAC. In this study, the significance of using Plasma Thromboplastin Cell Block (PTCB) routinely as an addition to CS in FNAC of palpable breast lesions. Aim: To determine the significance of PTCB as an adjunct in addition to CS to diagnose breast lesions. Materials and Methods: The present prospective observational study was conducted in the Department of Pathology, Chettinad Hospital and Research Institute, Kelambakkam, Chennai, Tamil Nadu, India, between July 2021 and June 2022 on 30 samples of palpable breast lesions. From the fine needle aspirates, smears were prepared and stained with Leishman and Papanicolaou stains. The residual material in the hub was rinsed in saline. The plasma-thromboplastin method was used to prepare CB, and Haematoxylin and Eosin (H&E) sections were made. A point scoring system was used and findings were compared to histopathology. IHC markers namely Estrogen Receptor (ER), Progesterone Receptor (PR), Human Epidermal Growth Factor Receptor-2 (HER2), Proliferation marker Ki-67 was utilised wherever appropriate. The results were analysed using Statistical Package for the Social Sciences (SPSS) software version 21.0. Results: Out of total 30 subjects, majority (n=9, 30%) were in the age group of 41-50 years. The mean scores of CS {background (0.93±0.25), cellularity (1.7±0.55), morphology (1.7±0.47) and architecture (1.03±0.32)} and PTCB {background (1.77±0.43), cellularity (1.77±0.48), morphology (1.8±0.48) and architecture (1.5±0.57)} were compared using the point scoring system. Though the mean scores of all four parameters were higher in PTCB than in CS, the statistically significant difference was seen in background (p-value=0.001) and architecture categories (p-value=0.0001). The PTCB finding as a screening test for predicting histopathological diagnosis showed a sensitivity of 94.44%, specificity of 100%, Positive Predictive Value (PPV) of 100%, Negative Predictive Value (NPV) of 92.3%, and 96.67% accuracy. IHC staining was feasible in CB and findings were comparable to biopsy. Conclusion: The routine use of PTCB technique in FNAC of breast lesions, along with smears, will aid in IHC, reducing diagnostic pitfalls, thereby reducing misdiagnosis and invasive procedures, particularly in suspicious for malignancy cases, which can lead to inappropriate radical treatment causing physical and psychological stress to patients. [ABSTRACT FROM AUTHOR]

Background: Tobacco-induced mucosal changes have been identified in exfoliated cells. The morphology of the exfoliated cells depends on the nature of the changes taking place in the epithelial layer; conversely, alteration in cytological pattern may be attributed to the changes occurring in the epithelial layer. Applying this possibility, exfoliative cytological techniques have been applied to examine the effect of tobacco on the oral mucosa. Aims and Objectives: The study was conducted to study the alterations in the cellular diameter of buccal mucosal cells. Materials and Methods: In a case controlled study, 240 participants were divided into four groups according to their habit of consumption of tobacco leaves, tobacco with areca nut, participants with oral lesions, and participants without any such habit. The groups were, further, subdivided according to their age. Desquamated cells from the buccal mucosa of were collected and smeared on a slide. The cells were stained with Papanicolou (rapid kit). Fifty cells per slide were studied and photomicrographs were obtained at ×40 magnification. The cellular diameter was obtained using the Leica 1000 software. Results: The mean cell diameter was found to be highest in the group of normal subjects (49.51±7.31 micron), followed by the group of tobacco users (43.52±1.92 micron) and Gutkha users (39.05±3.57 micron). The mean cell diameter was minimum (24.97±3.13 micron) in the group of oral lesion patients. According to the ANOVA test, the difference in cell diameter among various groups is found to be highly significant (P<0.0001). Conclusion: Tobacco used in any form can induce changes in the buccal mucosa. In our study, mean cellular diameter decreased from normal-cells to cells affected by oral lesions. [ABSTRACT FROM AUTHOR]

Programmed death ligand-1 (PD-L1) immunostaining, which aids clinicians in decision-making on immunotherapy for non-small cell lung cancer (NSCLC) patients, is sometimes performed on cytological specimens. In this study, differences in cytology fixation and cell block (CB) processing between pathology laboratories were assessed, and the influence of these differences on interlaboratory variation in PD-L1 positivity was investigated. Questionnaires on cytology processing were sent to all Dutch laboratories. Information gathered from the responses was added to data on all Dutch NSCLC patients with a mention of PD-L1 testing in their cytopathology report from July 2017 to December 2018, retrieved from PALGA (the nationwide network and registry of histo- and cytopathology in the Netherlands). Case mix-adjusted PD-L1 positivity rates were determined for laboratories with known fixation and CB method. The influence of differences in cytology processing on interlaboratory variation in PD-L1 positivity was assessed by comparing positivity rates adjusted for differences in the variables fixative and CB method with positivity rates not adjusted for differences in these variables. Twenty-eight laboratories responded to the survey and reported 19 different combinations of fixation and CB method. Interlaboratory variation in PD-L1 positivity was assessed in 19 laboratories. Correcting for differences in the fixative and CB method resulted in a reduction (from eight (42.1%) to five (26.3%)) in the number of laboratories that differed significantly from the mean in PD-L1 positivity. Substantial variation in cytology fixation and CB processing methods was observed between Dutch pathology laboratories, which partially explains the existing considerable interlaboratory variation in PD-L1 positivity. [ABSTRACT FROM AUTHOR]

Objective: The Paris System for Reporting Urinary Cytology (TPS) is a well‐known urinary diagnostic model; however, occasional false‐positives are a problem. To address this issue, we developed an improved algorithm (IA), based on additional cytological features, for TPS diagnosis. Methods: Cytological features were evaluated in 29 hard‐to‐classify cases, including 22 malignant cases and seven benign cases, using image analysis. The optimal IA was determined using the area under the receiver operating characteristic curve as an index. Re‐evaluation was performed by applying measured values to the TPS and IA algorithms. Results: Using TPS, 12 of the 22 malignant cases were reassigned to a more appropriate category, and the remaining 10 malignant cases remained hard‐to‐classify. Two of the seven benign cases were classified as suspicious for high‐grade urothelial carcinoma, and the remaining five benign cases remained in the original category. The IA, which included nuclear area as a parameter, showed the same diagnostic sensitivity as TPS, and three of the seven benign cases were reassessed as negative. Thus, the positive and negative predictive values of the IA were higher than those of TPS (84.6% and 100% vs 75.9% and 0%). Conclusions: The newly developed IA is a practical algorithm with which to address the limitations of TPS and thus may contribute to improved diagnostic accuracy. [ABSTRACT FROM AUTHOR]

We present a simple nanopore-electroporation (NanoEP) platform for delivery of nucleic acids, functional protein, and Cas9 single-guide RNA ribonucleoproteins into both adherent and suspension cells with up to 80% delivery efficiency and >95% cell viability. Low-voltage electric pulses permeabilize a small area of cell membrane as a cell comes into close contact with the nanopores. The biomolecule cargo is then electrophoretically drawn into the cells through the nanopores. In addition to high-performance delivery with low cell toxicity, the NanoEP system does not require specialized buffers, expensive materials, complicated fabrication processes, or cell manipulation; it simply consists of a generic nanopore-embedded water-filter membrane and a low-voltage square-wave generator. Ultimately, the NanoEP platform offers an effective and flexible method for universal intracellular delivery.

ObjectiveThe objective of this study was to determine the performance characteristics of the Tush brush (TB) compared with a saline moistened Dacron swab (DS) as anal cytology sampling devices.Materials and methodsTB and DS anal cytology tests were randomly collected from 146 patients presenting for anal cytology. High-resolution anoscopy and biopsies were obtained as indicated. Sensitivity and specificity as well as rates of satisfactory specimens were determined for each method using the areas under the receiver operating characteristic curve (AUCROC) and McNemar's test, respectively. Perceived discomfort of each device was determined using a visual analog scale and compared using a paired t test.ResultsThe adjudicated AUCROC, sensitivity, and specificity were greater, but not significantly different, for the brush (0.63, 85.5, and 40.0, respectively) compared with the swab (0.50, 79.6, and 33.3, respectively) when the anal biopsy results were considered the criterion standard. In the 1 subject diagnosed with anal cancer, the swab cytology result was normal, but the brush result was abnormal. Specimen adequacy was 95.2% for the brush and 93.2% for the swab. Mean discomfort (visual analog scale) scores were swab 28.5 mm versus brush 35.6 mm (p = .0003) with both scores within the minimal to moderate discomfort range.ConclusionsAnal cytology AUCROC, sensitivity, and specificity in detecting anal neoplasia were greater using the TB when compared with the DS. A novel anal cytology sampling device designed specifically to increase the detection of anal neoplasia would be clinically beneficial.

Technologies that convert transient protein-protein interactions (PPIs) into stable expression of a reporter gene are useful for genetic selections, high-throughput screening, and multiplexing with omics technologies. We previously reported SPARK (Kim et al., 2017), a transcription factor that is activated by the coincidence of blue light and a PPI. Here, we report an improved, second-generation SPARK2 that incorporates a luciferase moiety to control the light-sensitive LOV domain. SPARK2 can be temporally gated by either external light or addition of a small-molecule luciferin, which causes luciferase to open LOV via proximity-dependent BRET. Furthermore, the nested 'AND' gate design of SPARK2-in which both protease recruitment to the membrane-anchored transcription factor and LOV domain opening are regulated by the PPI of interest-yields a lower-background system and improved PPI specificity. We apply SPARK2 to high-throughput screening for GPCR agonists and for the detection of trans-cellular contacts, all with versatile transcriptional readout.

Primordial germ cells develop into oocytes and sperm cells. These cells are useful resources in reproductive biology and regenerative medicine. The mesenchymal stem cells (MSCs) have been examined for in vitro production of primordial germ cell-like cells. This study aimed to summarize the existing protocols for MSCs differentiation into primordial germ cell-like cells (PGLCs). In the limited identified studies, various models of mesenchymal stem cells, including those derived from adipose tissue, bone marrow, and Wharton's jelly, have been successfully differentiated into primordial germ cell-like cells. Although the protocols of specification induction are basically very similar, they have been adjusted to the mesenchymal cell type and the species of origin. The availability of MSCs has made it possible to customize conditions for their differentiation into primordial germ cell-like cells in several models, including humans. Refining germ cell-related signaling pathways during induced differentiation of MSCs will help define extension to the protocols for primordial germ cell-like cells production.

China has the considerable disease burden of cervical cancer, with the mortality and morbidity of cervical cancer showing an increasing and younger trend. Facing to the critical situation of cervical cancer control, it is urgent to explore the new methods that suitable for different resource areas for the early detection and treatment of cervical cancer. Recently, great progress has been made in the field of AI image classification, and scientists have developed many algorithms to identify cervical lesions and conducted corresponding studies on their accuracy. Here, by reviewing the papers published at home and aboard, which studied the applications value of AI in cervical cytology screening, colposcopy examination, diagnosis and treatment of cervical cancer, we summarized and discussed the current progress and challenges for AI's application in the area of cervical cancer control, in order to provide solid evidence for the future use of AI in improving human health.

Introduction: Despite the fact that Fine Needle Aspiration Cytology (FNAC) has been widely utilised in the preoperative diagnosis of breast lumps, the Conventional Smears (CS) have drawbacks, including difficulty in understanding the pattern or architecture of the lesion, determining invasiveness, Immunohistochemistry (IHC), false positives, and false negatives. Cytologists advise using Cell Blocks (CB) to increase the diagnostic precision of FNAC. In this study, the significance of using Plasma Thromboplastin Cell Block (PTCB) routinely as an addition to CS in FNAC of palpable breast lesions. Aim: To determine the significance of PTCB as an adjunct in addition to CS to diagnose breast lesions. Materials and Methods: The present prospective observational study was conducted in the Department of Pathology, Chettinad Hospital and Research Institute, Kelambakkam, Chennai, Tamil Nadu, India, between July 2021 and June 2022 on 30 samples of palpable breast lesions. From the fine needle aspirates, smears were prepared and stained with Leishman and Papanicolaou stains. The residual material in the hub was rinsed in saline. The plasma-thromboplastin method was used to prepare CB, and Haematoxylin and Eosin (H&E) sections were made. A point scoring system was used and findings were compared to histopathology. IHC markers namely Estrogen Receptor (ER), Progesterone Receptor (PR), Human Epidermal Growth Factor Receptor-2 (HER2), Proliferation marker Ki-67 was utilised wherever appropriate. The results were analysed using Statistical Package for the Social Sciences (SPSS) software version 21.0. Results: Out of total 30 subjects, majority (n=9, 30%) were in the age group of 41-50 years. The mean scores of CS {background (0.93±0.25), cellularity (1.7±0.55), morphology (1.7±0.47) and architecture (1.03±0.32)} and PTCB {background (1.77±0.43), cellularity (1.77±0.48), morphology (1.8±0.48) and architecture (1.5±0.57)} were compared using the point scoring system. Though the mean scores of all four parameters were higher in PTCB than in CS, the statistically significant difference was seen in background (p-value=0.001) and architecture categories (p-value=0.0001). The PTCB finding as a screening test for predicting histopathological diagnosis showed a sensitivity of 94.44%, specificity of 100%, Positive Predictive Value (PPV) of 100%, Negative Predictive Value (NPV) of 92.3%, and 96.67% accuracy. IHC staining was feasible in CB and findings were comparable to biopsy. Conclusion: The routine use of PTCB technique in FNAC of breast lesions, along with smears, will aid in IHC, reducing diagnostic pitfalls, thereby reducing misdiagnosis and invasive procedures, particularly in suspicious for malignancy cases, which can lead to inappropriate radical treatment causing physical and psychological stress to patients.

Objectives: The aims of this study was to investigate the use of cytologic samples for subclassification of lung adenocarcinoma and the cytologic-histologic correlation in lung adenocarcinoma subtypes using small samples. Methods and Methods: Cytological characteristics of lung adenocarcinoma subtypes were summarized by a literature review. Cytology samples from 115 patients with lung adenocarcinoma confirmed by small biopsies were classified by subtype. The diagnostic concordance of subtypes between biopsy and cytology samples was assessed. Results: Among the 115 cases, 62 (53.9%) had acinar predominant pattern, 16 (13.9%) were papillary predominant pattern, 29 (25.2%) had solid predominant pattern, 3 (2.6%) had lepidic predominant pattern, and 5 (4.3%) had micropapillary predominant pattern. All corresponding cytologic samples were classified into five subtypes based on cytomorphology features, with concordance rates of 74.2% (46 patients) in c-acinar subtype, 56.3% (nine patients) in c-papillary subtype, 24.1% (seven patients) in c-solid subtype, 66.7% (two patients) in c-lepidic subtype, and 40% (two patients) in c-micropapillary subtype. Collectively, the cytology and small biopsy concordance rate was approximately 57.4%. Conclusion: Subtyping of lung adenocarcinoma using cytologic specimens is challenging and the consistency rate varies with the subtype. Acinar predominant tumors have an excellent cytologic-histologic correlation compared to tumors with predominant solid or micropapillary pattern. Evaluating cytomorphologic features of different lung adenocarcinoma subtypes can reduce the false-negative rate of lung adenocarcinoma, particularly for the mild, atypical micropapillary subtype, and improve diagnostic accuracy. [ABSTRACT FROM AUTHOR]

The objective of this study was to verify the accuracy of the quantitative PCR (qPCR) method for in vivo detection of Macrorhabdus ornithogaster. A total of 100 faecal and gastric mucosa samples from avian cadavers were investigated, using cytological and qPCR techniques (budgerigars, Fischer's lovebirds, red-crowned parakeets, scarlet-chested parrots, eastern rosellas, domestic canaries, zebra finches, white Java sparrow). Using qPCR, the probability of detecting positive samples of droppings was significantly higher than in the faecal smear microscopy (P < 0.01). Cytology detected the presence of Macrorhabdus ornithogaster in 34 faecal samples, whereas qPCR detected 54 positive samples. In all 46 qPCR negative faecal samples, gastric smear qPCR was performed and also yielded negative results. Gastric smear qPCR was also performed in 20 cadavers where faecal qPCR has detected the presence of Macrorhabdus ornithogaster and in all samples confirmed the positive result. This verifies the accuracy of faecal sample qPCR for intravital diagnostics. Overall, the faecal qPCR technique appears to be extremely reliable, as it made it possible to detect all infected individuals, including those with negative stool or gastric cytology. [ABSTRACT FROM AUTHOR]