Fluorescence in situ hybridization (FISH) is a cytological method used to visualize specific oligonucleotide sequences within the cell. This method relies on the specific binding of a fluorescence-tagged probe, a short stretch of single-stranded polynucleotide, to its complementary sequence in the DNA or RNA, forming stable double-stranded hybrids. Fluorochromes, such as fluorescein, Alexa Fluor, cyanine dyes, or rhodamine, are attached to these probes to help in detecting their presence within the cell. Based on sequence complementarity, FISH allows for the visualization of the DNA or RNA with which they have hybridized. The distribution of these fluorochrome-tagged probes can be observed under a fluorescence or confocal microscope. The oligo(dT) FISH technique specifically utilizes a fluorochrome-tagged stretch of 40–50 thymidine (T) oligonucleotides that binds to the poly(A) tails of mature mRNAs within the cell. Newly transcribing pre-mRNAs and certain non-coding RNAs may not have poly(A) tails and therefore cannot be detected by this method. This step-by-step protocol outlines the oligo(dT) FISH technique for visualizing the cellular distribution of polyadenylated mRNAs in the tissues of Drosophila and other related model organisms.Key features• Oligo (dT) fluorescence in situ hybridization is a robust cytological technique to visualize polyadenylated RNAs in the cell.• This oligo(dT) FISH protocol is applicable for a wide range of animals and cell culture model systems.• This protocol is particularly useful to visualize mRNAs in thin tissues with a few layers of cells.

Background: Bronchial biopsy remains the reference standard for diagnosing pulmonary pathology; however, minimally invasive cytological techniques are increasingly used in clinical practice. This study aimed to determine the diagnostic accuracy of sputum cytology, bronchoalveolar lavage (BAL) cytology, transbronchial needle aspiration (TBNA), and bronchial brush cytology in detecting malignant pulmonary lesions using bronchial biopsy as the reference standard, and to compare their relative diagnostic performance.Methods: A prospective observational study was conducted at Government Medical College, Datia, from March to September 2025. A total of 120 adult patients with clinical and radiological suspicion of pulmonary pathology underwent sputum cytology and bronchoscopy-guided sampling (BAL, TBNA, brush cytology), followed by bronchial biopsy. Histopathology served as the reference standard. Diagnostic performance parameters were calculated. Suspicious cytology results were considered positive for malignancy for sensitivity analysis.Results: Of 120 patients, 77 (64.2%) had malignant lesions on biopsy. TBNA cytology demonstrated the highest sensitivity (92.2%) and excellent diagnostic accuracy (area under the curve (AUC) 0.969), followed by brush cytology (sensitivity 85.7%, AUC 0.974). BAL cytology showed moderate sensitivity (71.4%, AUC 0.887), while sputum cytology had lower sensitivity (49.4%, AUC 0.808). All modalities showed statistically significant associations with histopathology (p < 0.001). TBNA and brush cytology exhibited superior concordance with biopsy findings compared with BAL and sputum cytology.Conclusion: Bronchoscopy-guided cytological techniques, particularly TBNA and brush cytology, demonstrate high diagnostic accuracy and strong concordance with bronchial biopsy findings. These modalities serve as valuable complementary diagnostic tools in selected clinical settings but do not replace histopathological confirmation.

Pancreatic cancer is a highly lethal disease characterized by rapid onset, aggressive progression, and limited treatment options. The involvement of FoxM1 in the TGF-β/Smad signaling pathway has been linked to pancreatic cancer progression; however, the mechanisms behind the cooperative regulation of TGF-β signaling by FoxM1 and Smad4 remain poorly understood. In this study, we utilized molecular cytology techniques, animal models, and human pancreatic cancer tissues to investigate the role of FoxM1 in Smad4 stabilization and its regulation of TGF-β signaling. Our findings reveal that FoxM1 inhibits ubiquitin-proteasome-mediated degradation of Smad4, resulting in its stabilization. Once translocated into the nucleus, Smad4 binds to the FoxM1 promoter region, inducing FoxM1 expression and forming a positive feedback loop. Furthermore, we observed significantly higher expression of this feedback loop in pancreatic cancer tissues compared to adjacent normal tissues, with markedly elevated levels in poorly differentiated tissues compared to well-differentiated ones. Therefore, the loop aberrantly activates the TGF-β pathway, driving pancreatic cancer progression. These findings uncover a novel mechanism of TGF-β pathway activation and provide potential new targets for the prevention and treatment of pancreatic cancer.This study elucidates that FoxM1 functions to impede the ubiquitin proteasome-mediated degradation of Smad4, consequently stabilizing it. Following nuclear translocation, Smad4 binds to the FoxM1 promoter region, initiating FoxM1 expression and establishing a positive feedback loop. This loop plays a pivotal role in promoting pancreatic cancer development and migration by aberrantly activating the TGF-β pathway.

Abstract Veterinary diagnostic techniques are pivotal for the early identification and effective treatment planning, making them central to advancing veterinary oncology. They underpin the accurate identification and characterisation of neoplastic diseases, helping to guide effective treatment planning and improve animal health outcomes. This review emphasises the critical role of diverse diagnostic techniques, including cytology, histopathology, immunohistochemistry, electron microscopy, radiographic imaging (X-ray, computed tomography, positron mission tomography, magnetic resonance imaging), serology, and molecular techniques such as polymerase chain reaction (PCR), fluorescence in situ hybridization, and gene sequencing, that are critical for tumour diagnosis in veterinary practice. Despite their indispensability, these methods face significant challenges, mainly a lack of comprehensive standardisation and limited validation of established protocols and grading schemes. Recognising these related issues will assist in resolving them, which is vital for improving diagnostic accuracy, promoting innovation, and equipping veterinary professionals to make informed and effective clinical oncology decisions, thereby advancing both veterinary care and research.

Fine-needle aspiration cytology (FNAC) is an essential tool for evaluating canine mammary tumors (CMTs), yet its accuracy for grading requires validation. This study aimed to evaluate the diagnostic accuracy, malignancy grading, and architectural patterns using FNAC and agarose cell block (CBA) compared to histopathology in 30 CMTs obtained from surgical specimens. Additionally, the correlation between cytological grading and sentinel lymph node metastasis was investigated. Diagnostic efficacy for malignancy was 90% for FNAC and 97% for CBA. Regarding malignancy grading, concordance with histopathology was 65% for FNAC and 95% for CBA. Moreover, CBA allowed for morphological classification, showing moderate agreement (60%; k=0.50) with histopathological subtypes. A significant positive correlation (p=0.016) was observed between FNAC malignancy grade and inguinal lymph node metastasis. In conclusion, CBA proves to be a promising tool for tumor grading and architectural assessment, while the proposed cytological grading system serves as a feasible prognostic indicator for metastatic risk, highlighting the need for future studies to validate these findings in clinical practice.

Meiosis assures formation of both male and female gametes. However, cytological studies of female meiotic chromosome behavior in plants are scarce, mainly due to the comparatively low number and inaccessibility of female meiotic cells. We present Female Meiotic cell IDentification (FeM-ID), an approach for cytological identification of female meiotic cells in Arabidopsis. By employing ASY1-eYFP-TurboID, female meiotic cells (chromatin and cytoplasm) are specifically labeled by biotin. This facilitates their rapid microscopic identification and detailed assessment of female meiotic chromosome behavior. ASY1-eYFP-TurboID can be transformed or introgressed into different genetic backgrounds without any adverse effects on meiosis in either sex. Our approach enabled us to analyze female meiotic chromosome behavior, including bivalent and univalent frequencies, minimum chiasma numbers and chromosome segregation, in various ecotypes (Col-0, Ler-0, and Ws-2), colchicine-induced tetraploids (Col-4x) and selected meiotic mutants (sporulation 11-1 (spo11-1), meiotic recombination 11 (mre11), and mutL homolog 3 (mlh3), facilitating cytological studies of heterochiasmy (sex-specific differences in recombination rates or patterns). FeM-ID is compatible with other cytological techniques, such as immunolocalization and fluorescence in situ hybridization, which allowed us, for instance, to assess homologous chromosome pairing or to track individual chromosomes in female meiotic cells. In conclusion, FeM-ID represents an approach that fills a long-standing gap in the dissection of female meiotic chromosome behavior in Arabidopsis and holds the potential for application in other plant species.

Background and study aimsIntraoperative peritoneal lavage cytology (CY) influences prognosis in pancreatic cancer, and positive findings may lead to aborted surgery. Staging laparoscopy is traditionally used when no distant metastases are evident. We developed a novel method to assess CY using endoscopic ultrasound (EUS) and conducted a Phase 1 trial to evaluate its safety.Patients and methodsThis non-randomized, prospective Phase I1trial was conducted in three stages with safety monitoring after each stage. The study was approved by the institutional review board (2023–0-239) and registered (UMIN000052528). Nine patients with pancreatic ductal adenocarcinoma who were considering surgery were enrolled between September 2023 and August 2024. A 3F sheath was inserted into the upper abdomen under endoscopic and fluoroscopic guidance, followed by injection of 200 to 300 mL of saline. After postural adjustments and abdominal massage, transrectal EUS-fine-needle aspiration was used to aspirate pelvic fluid.ResultsMedian age was 68 years (range, 36–80); 77.8% were male. Resectability status: resectable/borderline/unresectable (considering conversion): 4/2/3. The procedure was successful in all cases, with a median aspirated volume of 32 mL (range, 10–125). No adverse events occurred, and all patients were discharged the next day.ConclusionsThis novel EUS-guided lavage cytology method was safe and feasible. A Phase 2 trial is planned.

Recent Advances in Cancer Diagnostics: Exotic Animal Medicine and Beyond.

Large type 3 (≥ 8cm) and type 4 gastric cancers are associated with extremely poor prognoses. The phase III JCOG0501 trial, which evaluated neoadjuvant S-1 plus cisplatin, failed to demonstrate any survival benefits. Docetaxel, oxaliplatin, and S-1 (DOS) have been explored as more effective perioperative regimens for these tumors. Eligible patients had large type 3 or type 4 gastric cancer without distant metastases, except for positive peritoneal cytology (CY). Patients received three cycles of neoadjuvant DOS (docetaxel 40mg/m2, oxaliplatin 100mg/m2, and oral S-1 at 80mg/m2/day for 14days), followed by gastrectomy with ≥ D2 lymphadenectomy and one year of adjuvant docetaxel plus S-1. The primary endpoint was the 3year progression-free survival (PFS) rate, with an expected value of 60% and a threshold of 45%. A one-sample log-rank test was performed with an α level of 0.10. Of the 48 patients enrolled, 27 had type 4 tumors (56.2%), and 10 (20.8%) had CY1. Overall, 91.7% of patients completed neoadjuvant DOS. R0 resection rate was achieved in 89.6% of patients, a pathological response grade ≥ 1b in 66.7%, and negative CY conversion in 80.0%. The 3-year PFS rate was 37.5% (95% confidence interval [CI], 24.1-50.6%; 80% CI 28.6-46.4%; p = 0.960), and the 3-year overall survival rate was 52.1% (95% CI 37.2-65.0%). Although neoadjuvant DOS therapy demonstrated favorable pathological responses, the 3-year PFS did not exceed the predefined threshold, and a survival benefit was not demonstrated.

366 Background: Large type 3 (≥8 cm) and type 4 gastric cancers are associated with extremely poor prognoses. Neoadjuvant chemotherapy (NAC) has been proposed as a potential strategy to improve outcomes in these patients. The phase III JCOG0501 trial, which compared neoadjuvant S-1 plus cisplatin followed by D2 gastrectomy with upfront surgery, failed to demonstrate a survival benefit for the neoadjuvant approach. Based on the results of the PRODIGY study, DOS therapy was considered a potential candidate for large type 3 or type 4 gastric cancer. Methods: Patients with large type 3 or type 4 gastric cancer without distant metastases, except for positive peritoneal cytology (CY), were eligible. Staging laparoscopy was mandatory. Participants received three cycles of neoadjuvant DOS therapy (docetaxel 40 mg/m² and oxaliplatin 100 mg/m² intravenously on day 1, and oral S-1 at 80 mg/m² for 14 days, repeated every 3 weeks), followed by gastrectomy with ≥D2 lymphadenectomy. Patients who achieved R0 resection subsequently received adjuvant docetaxel plus S-1 therapy for one year. The primary endpoint was the 3-year progression-free survival (PFS) rate, with an expected value of 60% and a threshold of 45%. A one-sided log-rank test was used with α=0.10 and power (1−β) =0.8. Results: Between October 2019 and February 2022, 48 patients were enrolled. The median age was 66 years (range: 44–79), and 29 patients (60.4%) were male. Twenty-seven patients (56.2%) had type 4 tumors. Positive CY was observed in 10 patients (20.8%). Clinical stage III and IV disease were observed in 24 (50.0%) and 11 (22.9%) patients, respectively. NAC was completed in 91.7% of patients. The 3-year PFS rate was 37.5% (95% confidence interval [CI]: 24.1–50.6%, 80% CI: 28.6–46.4%, p = 0.96), and the 3-year overall survival (OS) rate was 52.1% (95% CI: 37.2–65.0%). Among 10 patients with measurable lesions, the objective response rate was 50.0%. The R0 resection rate was 89.6%. A pathological response of grade 1b or higher was achieved in 66.7% of patients. CY conversion to negative was achieved in 80.0% of cases. Grade 3 or 4 adverse events during NAC, including neutropenia and appetite loss, occurred in 37.5% of patients. Conclusions: This study demonstrated a favorable pathological response and acceptable safety profile of neoadjuvant DOS therapy for large type 3 or type 4 gastric cancer. However, the 3-year PFS did not exceed the null hypothesis threshold, and the survival benefit of DOS therapy could not be demonstrated. Clinical trial information: CRB5180012 .

Crinum asiaticum L. is an important reservoir of phytocompounds containing galanthamine, lycorine, tazettine and others with diverse pharmacological uses. Due to high commercial demand for these promising compounds in pharmaceutical sector, an efficient in vitro micropropagation protocol optimization study was conducted via direct somatic embryogenesis in C. asiaticum. The regenerated plants were subject to genetic fidelity assessment; and the phytochemical composition was analysed and compared with donor plants. In this investigation, the bulb-scales were used as explants onto media containing different PGRs for various regeneration processes. In media containing BAP and NAA, somatic embryos were formed directly on bulb-scale explant surfaces with the highest (95.83%) being at MS medium + 2.7 µM NAA + 4.4 µM BAP. The occurrence of somatic embryos at different stages was confirmed by histological and scanning electron microscopic (SEM) analysis. The embryos were later converted to shoots on 2.2-8.8 µM BAP augmented MS medium, with highest germination percentage of 75 ± 7.22 at 4.4 µM BAP. These regenerated plants were successfully transferred to medium containing NAA, IBA or IAA for rooting and the best rooting response (91.67% rooting frequency, 7.67 mean root numbers/shoot and 7.5 ± 0.6cm average root length) was noted at 5.4 µM NAA. The plants were transferred to greenhouse with pretty good growth and survival. The genetic fidelity of tissue cultured plants was checked through cytological, flow cytometric and SCoT marker-based PCR technique. The root tips of in vitro raised and mother plants showed 2n = 44 chromosome numbers, and the flow cytometric histograms revealed similar fluorescence peaks with nuclear 2C DNA content of 31.79 and 31.51pg, respectively, displaying no change in ploidy level. Six SCoT primers based genetic homogeneity study showed 42 scorable, monomorphic bands, confirming true-to-type regenerated plants. Finally, the GC-MS based metabolite profiling of in vivo and in vitro raised plants were conducted, which exhibited a wide range of bioactive compounds like tazettine, squalene, gamma-tocopherol, beta-sitosterol, glycidyl palmitate, glycidyl oleate of pharmacological significance. The current study presents an effective method for genetically stable clonal propagation of C. asiaticum for extraction of compounds like tazettine, squalene, beta-sitosterol for pharmaceutical applications.

Our group revealed that the combination of intra-operative stamp cytology and peritoneal lavage cytology (CY) improved the identification of individuals with high risk of peritoneal metastasis. In this exploratory Phase II study, we aimed to evaluate the effect on relapse-free survival (RFS) of extensive intraoperative peritoneal lavage (EIPL) for gastric cancer with positive peritoneal cytology (CY1) and/or stamp cytology positive (stamp+). This study was a single arm, multi-institutional, exploratory phase 2 trial to assess the effects of EIPL after open gastrectomy for gastric cancer with CY1 and/ or stamp+. The primary endpoint was RFS. Secondary endpoints were overall survival (OS), postoperative recurrence site and incidence of postoperative adverse events. Between 2017 and 2021, 13 patients from 2 institutions were enrolled in this study. Because of the recent decline in open abdominal surgery, the number of cases did not increase and the trial was closed due to lack of applicants at 13 cases. Median 3-year RFS was 14.5 months (95% CI 5.4-NA), median 3-year OS was not reached (95% CI 14.5-NA) and median3-year peritoneal RFS was 16.0 months (95% CI 5.4-NA). Median 3-year peritoneal RFS rate was 83% in CY0 and stamp+ cases (n=6), and 0% in CY1 and stamp+/- cases (n=7). (Log-rank p=0.015). Because of the slow accrual pace and early stop of the trial, we were not able to evaluate the prespecified endpoints thoroughly. However, EIPL might be effective to prevent perineal recurrence, especially in CY0 and stamp+ case.

Background:Peritoneal washing (PW) cytology has become a widely accepted procedure for staging of gynecological malignancies.Aims:To compare conventional cytology, liquid-based cytology (LBC) and cell block in the evaluation of PWs.Materials and Methods:A total of 50 clinically suspected cases of gynecological malignancies were enrolled, which included 41 ovarian neoplasms, eight uterine, and one case of cervical malignancy. The age of the women ranged from 13 to 74 years. The PW samples obtained in all cases were subjected to conventional cytology, LBC, and cell block preparations. Immunohistochemistry (IHC) was applied on cell blocks and analyzed. A comparative evaluation and statistical correlation between the three techniques was performed.Results:The number of cases reported as positive for malignancy on conventional smear and LBC were 9 (18%) and 11 (22%), respectively. Fifteen (30%) cases were reported as positive for malignancy on cell block after IHC analysis. All three cytological techniques showed a good correlation in diagnosing peritoneal metastasis on evaluation by Cohen’s kappa score (≤ 1.00). However, the overall sensitivity, specificity, and diagnostic accuracy of PW cytology was 43.8%, 94.4%, and 62%, respectively, taking histopathology as the gold standard. The most common malignancy which led to peritoneal fluid involvement was high-grade serous carcinoma of the ovary.Conclusion:The study demonstrates the usefulness of combining the three cytological techniques to diagnose peritoneal metastasis in gynecological malignancies. Cell block with IHC can help in reducing the false-negative cases and further adds to objectivity in ambiguous cases.

Background: Serous effusions are the fluids accumulated in body cavities like the pleural, pericardial, and peritoneal cavities. Detection of malignant cells in these effusions helps in staging and planning the course of management. Accurate identification of the malignant cells using conventional cytological examination is a diagnostic challenge. Argyrophilic nucleolar organiser regions (AgNORs) are a simple cytological technique that is useful for detecting these malignant cells in effusions. Hence, in the present study, the silver staining method is applied to differentiate malignant cells from reactive mesothelial cells in serous effusions. The objective is to determine the significance of AgNOR staining in differentiating benign and malignant serous effusions and to determine the prevalence of malignant changes in serous effusions. Material and Methods: This is a sectional study conducted from August 2022 to December 2023 in the Department of Pathology at Basveshwara Medical College and Hospital. This study includes serous effusions sent to the central lab for cytological examination. All samples were centrifuged, and smears were prepared and stained with H&amp;E and AgNOR. AgNORs are counted as black dots in the nuclei of 100 cells using 100x oil immersion. The pattern of AgNOR dispersion and its shape will be compared in reactive and malignant cases. Results: Out of 310 samples, 176 were ascitic, and 134 were pleural fluids. Prevalence of malignancy in these samples was 11.95%. Mean agnor count in NFM was 2.33 +/- 0.67, AUS was 5.68 +/- 1.99, SFM was 7.87 +/- 2.25, and in malignancy was 11.04 +/- 0.9, indicating that the mean count is higher in malignant cells than in benign cells. Benign cells had Agnor dispersion of 0-1+, and malignant cells had dispersion of 2+- 3+. Similarly, benign cells had an average size of 0-1+ and malignant cells had an average size of 2+-3+. Agnor size had a sensitivity of 91.89%and a specificity of 97.76% in detecting malignant cells. Agnor dispersion had a sensitivity of 94.59%and a specificity of 99.1% in detecting malignant cells. Conclusion: Mean Agnor count is significantly elevated in malignant cells when compared to benign cells. Along with increased size and dispersion of Agnor, a count is seen in malignant cells of serous effusion. Agnor stain is a particularly useful and rapid diagnostic test for differentiating benign from malignant cells when routine cytology fails. Keywords: AgNor staining, serous effusions: Agnor size, Agnor dispersion.

Mycetoma is a chronic and neglected inflammatory disease prevalent in tropical and subtropical regions. It can lead to severe disability and social stigma. The disease is classified into two types based on the causative microorganisms: eumycetoma (fungal) and actinomycetoma (bacterial). Effective treatment strategies depend on accurately identifying the causative agents. Current identification methods include molecular, cytological, and histopathological techniques, as well as grain culturing. Among these, histopathological techniques are considered optimal for use in endemic areas, but they require expert pathologists for accurate identification, which can be challenging in rural areas lacking such expertise. The advent of digital pathology and automated image analysis algorithms offers a potential solution. This report introduces a novel dataset designed for the automated detection and classification of mycetoma using histopathological images. It includes the first database of microscopic images of mycetoma tissue, detailing the entire pipeline from species distribution and patient sampling to acquisition protocols through histological procedures. The dataset consists of images from 142 patients, totalling 864 images, each annotated with binary masks indicating the presence of grains, facilitating both detection and segmentation tasks.

Mitosis is a fundamental process in plant growth and development because it governs the production of new cells in meristematic tissues. Examining its stages provides insights into genetic stability and cellular behavior, which are relevant to agriculture, crop improvement, and environmental monitoring. Although cytological studies using Allium species are well established, there is continued value in demonstrating how classical staining methods, such as acetocarmine, effectively visualize mitotic events in educational and basic laboratory settings. This study aimed to identify and describe the stages of mitosis in onion (Allium cepa) and spring onion (Allium fistulosum) root tips using acetocarmine-stained squash preparations. Fresh root tips were hydrolyzed in hydrochloric acid, stained with acetocarmine, and examined under a compound light microscope. The observed mitotic stages were documented based on chromatin morphology and nuclear characteristics. Both species exhibited similar observable mitotic features under the staining and microscopic conditions used, with clear visualization of prophase, metaphase, and telophase, identified by chromatin condensation, chromosome alignment, and nuclear reformation, respectively. Anaphase was not observed, likely due to its brief duration relative to the other phases. Overall, the results demonstrate that acetocarmine staining provides reliable visualization of major mitotic stages, although it lacks sufficient resolution to distinguish individual chromosomes. These findings confirm that simple cytological techniques remain effective tools for demonstrating mitosis in instructional and introductory research contexts. Future studies may employ DNA specific stains, digital imaging, and larger sample sizes to capture short-lived stages and enable quantitative analysis.

Accurate brain tumor diagnosis guides effective management, with large excision biopsy as the gold standard. Intra-operative cytologic smear techniques offer rapid analysis, particularly useful in resource-limited settings. The aim of this study was to evaluate the diagnostic accuracy and clinical utility of intra-operative cytologic smear techniques in brain tumor surgery at the University Teaching Hospital of Kigali (CHUK). To evaluate the diagnostic accuracy and clinical utility of intra-operative cytologic smear compared to large excision biopsy in brain tumor surgery at CHUK. A prospective cross-sectional study involving 77 patients with radiologically confirmed brain tumors undergoing surgery at CHUK. Squash cytology diagnoses were compared with final histopathology diagnoses for concordance. Squash cytology showed 89.6% agreement with histopathology. Major tumor types such as gliomas and meningiomas were reliably identified, facilitating timely surgical decisions. Intra-operative squash cytology is a rapid and reliable adjunct to large biopsy, improving brain tumor management in low-resource settings. Integration of advanced techniques could enhance diagnostic precision further.

Sporotrichosis is a subcutaneous and systemic mycotic infection affecting humans and animals. It is caused by a pathogenic dimorphic fungus of the genus Sporothrix. This study aimed to identify the clinical signs, prevalence, and risk factors of sporotrichosis in pet cats in Denpasar city. The sample of this study was the skin surface of cats, with a total sample of 101 cats. This study was conducted from November 2024 to January 2025. The examination method was carried out in four ways: physical examination, acetate tape preparation, and impression smear. The collection of skin surface samples was then examined by cytological techniques (Diff-Quik Staining). The results of this study showed that the clinical signs of one sporotrichosis-positive female cat included pruritus with a score of 4/10 in the head, face, and ear areas; the texture of the skin surface was rather rough and dry; there were crusts in the area between the right and left eyes; there was a lot of squama on most of her body's skin; and there was purulent discharge in both nostrils. The prevalence of sporotrichosis in pet cats in Denpasar city was 1%. Statistical analysis showed that factors such as sex, age, breed, husbandry, presence or absence of skin wounds, bathing frequency, and sterilization status were not risk factors for sporotrichosis.

Cervical cancer is primarily associated with the presence of human papillomavirus (HPV), with high-risk genotypes HPV-16 and HPV-18 being the focus of vaccination programs in developing countries such as Chile. Preventive screening for cervical cancer in women aged 25 to 64 years remains centered on cytological techniques and is primarily performed based on clinical suspicion of cervical lesions. However, extended screening for HPV genotypes using DNA amplification methods is not routinely applied to the Chilean female population yet. This study aimed to determine the prevalence of high- and low-risk HPV genotypes in women without known risk factors in a city in northern Chile. Cervicovaginal brushing samples were obtained from 390 women from Antofagasta city, Northern Chile, aged between 25 and 64 years; genomic DNA was extracted, and multiplex real-time PCR analysis was used to identify a larger group of high- and low-risk HPV genotypes. Among 390 samples, HPV prevalence was 36.9%, of which 54.9% were high-risk genotypes, 18.7% were low-risk genotypes, and 26.4% showed mixed infection with both high- and low-risk genotypes. High-risk genotypes 16, 58, 39, and 31 were the most frequently identified among HPV-positive samples. Furthermore, a significant association was observed between HPV presence and both age and suspicion of cervical alteration, and women testing positive for other sexually transmitted infections (STIs) were more likely to acquire HPV. Implementing a screening program that incorporates extended HPV genotyping in Chile, including testing for high-risk genotypes 16, 18, 31, 39, and 58, is crucial to optimize control, early detection, and vaccination efforts for Chilean circulating HPV genotypes that are not covered by the actual vaccine, thus contributing to a more effective reduction in the burden of disease associated with the virus.

High-quality RNA is essential for accurate molecular testing. This study evaluates the impact of cytological preparation techniques (May-Grünwald-Giemsa [MGG], Papanicolaou [PAP], Diff-Quik, and air-dried) on RNA quality in smear slides. A total of 182 smears were prepared from fresh surgical specimens of 26 patients using seven different techniques. RNA was isolated, reverse-transcribed, and analyzed using quantitative polymerase chain reaction (qPCR). RNA quality was assessed using ΔCt (ΔCt=45 - Ct, cycle threshold), where higher ΔCt indicates better RNA quality. RNA quality, measured by ΔCt, showed clear differences (p<.001) in-between preparation methods, whereas RNA concentration did not differ significantly among smear types (p=.07). MGG-stained smears (both film- and coverslip-mounted) demonstrated the highest and most consistent ΔCt values. PAP-stained smears yielded the lowest ΔCt values, indicating the poorest RNA quality. Air-dried unstained smears showed highly variable ΔCt values and frequent amplification failures. Diff-Quik preparations had intermediate performance. Mounting method (film vs. coverslip) did not significantly affect RNA quality. Among cytology smear techniques, MGG provided the best RNA preservation, PAP the worst, and air-dried slides yielded inconsistent results. These findings highlight the critical role of smear preparation in preserving RNA for molecular testing, especially RNA-based next-generation sequencing.