Cap-Specific m6Am Methyltransferase PCIF1/CAPAM Regulates mRNA Stability of RAB23 and CNOT6 through the m6A Methyltransferase Activity.

Chemical modifications of cellular RNAs play key roles in gene expression and host defense. The cap-adjacent N6,2'-O-dimethyladenosine (m6Am) is a prevalent modification of vertebrate and viral mRNAs and is catalyzed by the newly discovered N6 methyltransferase PCIF1. However, its role in gene expression remains unclear due to conflicting reports on its effects on mRNA stability and translation. In this study, we investigated the impact of siRNA-mediated transient suppression of PCIF1 on global mRNA expression in HeLa cells. We identified a subset of differentially expressed genes (DEGs) that exhibited minimal overlap with previously reported DEGs. Subsequent validation revealed that PCIF1 positively and negatively regulates RAB23 and CNOT6 expression, respectively, at both the mRNA and protein levels. Mechanistic analyses demonstrated that PCIF1 regulates the stability of these target mRNAs rather than their transcription, and rescue experiments confirmed the requirement of PCIF1's methyltransferase activity for these regulations. Furthermore, MeRIP-qPCR analysis showed that PCIF1 suppression significantly reduced the m6A levels of RAB23 and CNOT6 mRNAs. These findings suggest that PCIF1 regulates the stability of specific mRNAs in opposite ways through m6A modification, providing new insights into the role of m6Am in the regulation of gene expression.

Preparation and application of multiple particle binding-liposomes for electrochemiluminescent signal amplification in bioassays.

In this study, multiple particle binding-liposomes (MPB-Lips), encapsulating the luminophore tris(2',2-bipyridyl)ruthenium (II) complex ([Ru(bpy)3]2+), were developed as an electrochemiluminescence (ECL) signal amplifier and were applied to detect the model analyte streptavidin (SA) using the indirect competitive ECL method. The MPB-Lips were prepared by mixing various ratios of two different liposomes-one containing a phospholipid with a primary amine group and a biotinyl group (BIO/NH2-Lip) and one containing a phospholipid with an N-hydroxysuccinimide group (NHS-Lip) to allow binding between particles via amide bonds. Quartz crystal microbalance analysis using SA-modified gold-coated quartz crystals showed that the frequency shift values of MPB-Lips gradually decreased in the order BIO/NH2-Lip:NHS-Lip = 1:0 < 1:1 < 1:3 < 1:5. This indicated that MPB-Lips were successfully formed. The indirect competitive ECL method using SA-modified gold electrodes showed that the 1:5-Lip system had greater sensitivity than the 1:0-Lip system-the limit of detection and quantification values for the systems were 1.84 and 6.30 µg mL-1 for 1:0-Lip, and 1.20 and 1.74 µg mL-1 for 1:5-Lip. Finally, the recovery of SA spiked in fetal bovine serum samples using the 1:5-Lip system showed good accuracy and precision with a recovery rate of 83-106% and relative standard deviation of 4-14%. Our study demonstrated that the MPB-Lips system was an effective and useful ECL amplifier and the ECL method using MPB-Lips could be applied to detect an analyte in a real sample.

Decline in the Conception Rate of Wild Japanese Monkeys after the Fukushima Daiichi Nuclear Power Plant Accident.

We examined the conception rate of wild Japanese monkeys (Macaca fuscata) in Fukushima City that were exposed to radiation as a result of the Fukushima Daiichi Nuclear Power Plant accident in March 2011. The conception rate in the year of delivery from 2009 to 2022 was estimated by dissecting individuals that were euthanized by the government for population control as a countermeasure against crop damage. To evaluate the effects of exposure, the cumulative exposure dose for each individual was calculated using the concentration of radiocesium deposited in the soil at the capture site and the concentration of radiocesium in muscle estimated from the aggregated transfer factor. There were no significant differences in conception rates across all age classes over time. In terms of conception rates by age class, there was a significant decrease post-exposure compared with pre-exposure in the age class ≥ 8 years, but no significant differences in the age class 5-7 years. The non-ovulation rate did not significantly differ between the pre- and post-exposure periods for any age class. Body fat index, which can affect fertility, was compared between the pre- and post-exposure periods, and no significant differences were found in either age class. In contrast, the median total cumulative exposure (cumulative internal exposure + cumulative external exposure) was significantly higher in the age class ≥ 8 years compared with the age class 5-7 years. These results suggest that the total cumulative exposure dose may be one of the reasons for the lower conception rate in the post-exposure period among the age class ≥ 8 years.

Non-Invasive Myocardial Work Indices in Infants Born to Mothers With Diabetes in Pregnancy.

BACKGROUND: This study used echocardiography to investigate non-invasive myocardial work (MCW) indices in infants born to mothers with diabetes mellitus (DM) in pregnancy (gestational DM: GDM), including those diagnosed under novel classification criteria and those with pre-existing DM.Methods and Results: This single-centered, retrospective study included 25 infants born to mothers with GDM (termed "infant with GDM"), which was diagnosed by oral glucose tolerance test results during pregnancy or the presence of diabetes before the current pregnancy. We evaluated the relationship between the infant's MCW, such as global constructive work (GCW), global work index (GWI), global work efficiency (GWE), and global wasted work (GWW), and the mother's GDM maximal HbA1c during pregnancy. HbA1c level in GDM significantly negatively correlated with GWI* (r=-0.565) and GCW* (r=-0.641). In infants with GDM, GWI and GCW were significantly higher with <6.5% HbA1c than in those with >6.5% HbA1c GDM; however, the specific-layer global longitudinal strain analyses did not show any significant differences between the groups. The pressure-strain loop in infants with >6.5% HbA1c in GDM tended to be smaller than in those infants with <6.5% HbA1c GDM. CONCLUSIONS: The hyperglycemic environment of GDM leads to impaired MCW in the infants. MCW is useful for screening for cardiac illnesses among infants with GDM. Appropriate maternal blood glucose management while maintaining HbA1c <6.5% might be beneficial for the cardiac performance of infants with GDM.

Clinical, cytological, and pathological characteristics of metastatic renal cell carcinoma to the thyroid: A study of 14 cases at a Japanese single institution.

A preoperative diagnosis of metastatic renal cell carcinoma to the thyroid (MRCCT) is important for determining clinical management but is challenging even in cases with a clinical history of renal cell carcinoma (RCC). This study aimed to elucidate the clinical, cytological, and pathological characteristics of MRCCT. Fourteen MRCCT cases extracted from 18 320 malignant thyroid tumors were included in this study. Twelve MRCCT (85.7%) occurred as solitary lesions and the most frequently suspected lesions on ultrasonography were follicular tumors. On cytology, 46.2% of cases were reported as RCC or suspected RCC; a medical history of RCC and immunocytochemistry were helpful in interpretation. RCC metastasized to a follicular adenoma in 50.0% of the solitary lesions. MRCCTs with a long interval from the initial presentation, solitary lesion, and Ki-67 labeling index <10% showed significantly longer disease-free survival. MRCCT is characterized by a long interval from the initial presentation of RCC, appearance as a solitary nodule, ultrasonographic similarity to follicular tumors, sharing cytological findings with primary thyroid tumors, and high frequency of metastasis within follicular adenoma. A long interval from the initial presentation, occurrence as a solitary lesion, and low Ki-67 labeling index may be favorable prognostic factors.

Medial patellar luxation grade IV according to the Singleton system can be classified into skeletally immature and skeletally matured types.

OBJECTIVE: To determine the signalment and musculoskeletal morphology of small-breed dogs affected by medial patellar luxation (MPL) grade IV based on the age of the CT scan. ANIMALS: 40 small-breed dogs (54 limbs) with MPL grade IV. PROCEDURES: Dogs that had undergone corrective surgery for MPL grade IV and had performed CT of the hind limb before surgery were included. Signalment (age, body weight, sex, laterality, and breed) and concomitant cranial cruciate ligament rupture (CrCLR) were recorded. Femoral inclination angle, anatomical lateral distal femoral angle (aLDFA), femoral torsion angle, quadriceps muscle length to femoral length ratio (QML/FL), and patellar ligament length to patellar length were obtained by CT images. The dogs were categorized into 2 groups based on their age at the time of the CT scan, the skeletally immature group and the skeletally matured group. Signalment and group were included in the multiple regression analysis to determine the factors associated with each measurement parameter. A logistic regression analysis was conducted to determine the risk of CrCL concomitant with age. RESULTS: The multiple regression model demonstrated that the group was associated with the value of aLDFA and QML/FL. aLDFA was higher, and QML/FL was lower in group SI than in group SM. CrCLR was present in 5/54 limbs (9.2%), with a mean age of 70.8 months and it was associated with increasing age. CLINICAL RELEVANCE: In Singleton's classification, dogs classified as grade IV can be categorized into 2 groups based on musculoskeletal morphology and pathophysiology: the skeletally immature and skeletally matured types.

Influence of radiation exposure to delayed fetal growth in wild Japanese monkeys after the Fukushima accident.

Wild Japanese monkeys (Macaca fuscata) were exposed to radiation after the Fukushima Daiichi nuclear accident in 2011. To clarify the biological effects of radiation exposure on their fetal growth, pregnant monkeys and their fetuses were analyzed. These animals were collected between 2008 and 2020 (before and after the accident in 2011) living in Fukushima City, approximately 70 km from the nuclear power plant. Multiple regression analyses were conducted with fetal body weight (FBW) and fetal head circumference (FHS) as objective variables, and maternal and fetal factors as explanatory variables. The maternal factors were relative exposure dose rate (REDR), age, body weight, body length, fat index, and parity. The fetal factors were crown ramp length (CRL) and sex. Multiple regression analyses showed that FBR and FHS growth were positively associated with CRL, maternal body length, and negatively associated with REDR. Since the relative growth of FBR and FHS to CRL decreased with increasing REDR, radiation exposure due to the nuclear accident may have contributed to the delayed fetal growth observed in Japanese monkeys.

X-ray analytic microscopy with CT-guided sample extraction to analyze potential projectile fragments in a dead serow.

In forensic pathology, it is important to detect and recover as evidence residual metal particles and projectiles when evaluating potential gunshot wounds. This process can be challenging when the bullets are fragmented. This report presents our experience using multiple modalities to analyze the wound of an illegally killed Japanese serow (Capricornis crispus) found in a mountainous region without its head and hind limbs. We performed postmortem computed tomography (PMCT) and necropsy that showed a distant-range gunshot wound of the neck likely created by a centerfire rifle. A postmortem cut through the neck and absent head precluded a complete evaluation. To determine the composition of the metal-like fragments in the neck, sampling and metal analysis were performed in two ways. Samples extracted from the exposed wound surface without CT guidance were analyzed directly using inductively coupled plasma mass spectrometry (ICP-MS). Samples from the bone, muscles, and cervical cord extracted under CT guidance were analyzed using x-ray analytic microscopy (XGT-9000, HORIBA, Kyoto, Japan). In wound surface samples, silicon and iron were detected as the main components on ICP-MS, suggesting that the materials were gravel. The samples taken under CT guidance and analyzed with XGT-9000 revealed that the high-density CT areas were mainly composed of lead. Combining CT-guided sample extraction and XGT-9000 can be useful for retrieving true projectile fragments and avoiding the confusion created by erroneously sampling non-projectile surface materials.

Long-term histological effects of high-dose prednisolone administration on the mitral valve in normal Beagle dogs.

Background: In recent years, left ventricular hypertrophy and cardiac dysfunction have been reported in human and canine patients with hypercortisolism and in dogs treated experimentally with high-dose prednisolone. However, to our knowledge, there have been no reports on the effects of hyperglucocorticism (HGC) on the mitral valve (MV). Aim: This study aimed to compare the MV in dogs treated with high-dose prednisolone with that in healthy dogs to investigate the effects of HGC on the MV. Methods: We investigated the effects of HGC on the MV by comparing samples obtained from high-dose glucocorticoid (GC)-treated (P) and healthy (C) dogs. The P group included healthy Beagle dogs (n = 6) treated with prednisolone (2 mg/kg, bid, po) for 84 days and the C group included healthy Beagle dogs (n = 6) euthanized for unrelated reasons. The anterior and posterior mitral leaflets (AML and PML, respectively) from both groups were harvested and stained with hematoxylin-eosin, Alcian blue, and Masson trichome. Additionally, adiponectin (ADN) and GC receptor immunohistochemistry were performed. Histological evaluation was performed in the atrialis, spongiosa, fibrosa, and all layers of the proximal, middle, and distal regions of the AML and PML. Results: The proportion of the spongiosa layer thickness to the total thickness was higher in the P than in the C group (proximal and middle AML). However, the proportion of the fibrosa layer thickness to the total thickness was lower in the P than in the C group (middle PML). Areas of acidic sulfated mucosubstance deposition were smaller in the fibrosa layer and all layers (middle AML), while those of collagen deposition were smaller in the spongiosa and total layers (proximal and middle AML), in the P than in the C group. Additionally, ADN expression in the spongiosa layer was higher in the P than in the C group (middle AML). Conclusion: These findings suggest that long-term administration of synthetic GCs induces histological changes in the MV. These changes may lead to MV dysfunction in dogs with HGC.

Time dependence of 137Cs contamination in wild Japanese monkeys after the Fukushima Daiichi nuclear accident.

Over the 10 years immediately after the Fukushima Daiichi nuclear accident, we measured the changes in the muscle 137Cs concentration (Bq/kg) of wild Japanese monkeys living in Fukushima City, which is located approximately 70 km from the Fukushima Daiichi Nuclear Power Plant. The muscle137Csconcentration, which was observed at a maximum of 13,500 Bq/kg immediately after the accident, had decreased to several hundred Bq/kg 10 years later. The muscle 137Cs concentration was significantly related to the soil contamination levels (10,000-30,000, 30,000-60,000, 60,000-100,000, and 100,000-300,000 Bq/m2), sex, age class (immature, mature), body weight (> 5000 g, 5000-10,000 g, < 10,000 g), and seasons (the cold period from December to April, the warm period from May to November).The value of muscle 137Cs concentration and the aggregated transfer factor (Tag: calculated by dividing muscle 137Cs concentration [Bq/kg] by soil 137Cs deposition density at the capture site [Bq/m2]) apparently decreased with time for several years. However, post hoc pairwise comparisons showed no difference from 2017 to 2020, and the accumulation of 137Cs in muscle may continue for some time.

The impact of coronavirus disease 2019 (COVID-19) in Japanese companion animal clinics.

The coronavirus disease 2019 (COVID-19) pandemic has had a tremendous impact on people's lives throughout the world. A cross-sectional study was conducted to clarify the influence of COVID-19 on Japanese companion animal clinics. A self-administered electronic questionnaire regarding the incidence of COVID-19, hygiene management, the influence on clinical service and employment, and mental stress of staff was conducted for workers in animal clinics between 1 May and 10 June 2021. Questions concerning the hygiene management before the occurrence of COVID-19, under the first state of emergency, and at the time of the survey were asked using the Likert scale. Kessler 6 (K6) was used as an indicator of mental distress. In total, 430 individuals responded. Of these, 4.9% experienced COVID-19 infection in staff. Hygiene management was strengthened, but no particular practice was statistically associated with the occurrence of COVID-19 cases in staff. In Nakaya's variation of Scheffe's paired comparison, the highest prioritized goal in the clinics was the prevention of nosocomial infection with COVID-19. The prevalence of serious mental illness (K6 >=13) was 11.1% (95% confidence interval: 6.3-18.6%). Multivariable negative binomial regression found four risk factors for psychological distress: veterinary nurse (P=0.016 with veterinarians and P<0.01 with other staff), female (P=0.004), fear of infection at work (P<0.001), and stress by refraining from going out (P<0.001). Directors of clinics are recommended to take care of female veterinary nurses for distress.

Lymphoma in Miniature Dachshunds: A retrospective multicenter study of 108 cases (2006-2018) in Japan.

BACKGROUND: Miniature Dachshunds (MD) are predisposed to lymphoma with disease onset of young age and long-term survival. OBJECTIVES: To compare clinical features and survival time of lymphoma in MD and non-MD. ANIMALS: One hundred and eight MDs with lymphoma and 149 non-MD breed dogs with lymphoma were included in the study. METHODS: This was a retrospective multicenter observational study. Lymphoma was classified based on signalment, histopathology/cytology, and anatomical site of the disease. For each type of lymphoma, median survival time was analyzed by Kaplan-Meier estimates and life table analysis. Prognostic factors for large-cell gastrointestinal lymphoma (LGIL) were analyzed using Cox regression. RESULTS: Gastrointestinal lymphomas were more common in MDs (53/108) compared to non-MDs (41/149). The multicentric lymphoma was most common in non-MD breed dogs (74/149) compared to MDs (33/108). The median age that dog developed lymphoma in MD and non-MD were both 10 years old; however, lymphomas were more frequently observed in younger dogs (<4 years) in MDs (20/108) compared to non-MDs (9/149; P = .002). Seventy percent were diagnosed with B-cell with median age of diagnosis was 3 (1-14) years. Mott cell differentiation was observed in 6 dogs. Age <4 years and B-cell phenotype were significant factors for longer survival time in MD with LGIL. CONCLUSIONS AND CLINICAL IMPORTANCE: Lymphomas in MDs involved gastrointestinal lesions at higher frequency compared to other dog breeds examined. B-cell lymphoma was more common in early-onset LGIL in MD and cases that involved Mott cell differentiation were observed. Awareness of this specific presentation of lymphoma in dogs will possibly affect the treatment decision process for the owners of MD with LGIL.

B-cell to T-cell ratio as a novel indicator in flow cytometry in the diagnosis of thyroid lymphoma.

Preoperative flow cytometry is recommended to prove the monoclonality and confirm the diagnosis of thyroid lymphoma. However, lymphoma cases without light chain restriction may also have monoclonality. The aim of our study was to identify a novel marker for thyroid lymphomas using aspirated materials for flow cytometry. We retrospectively analyzed 26 patients with primary thyroid lymphomas and 16 patients with benign lymphoproliferative lesions. The materials for flow cytometry were obtained by fine-needle aspiration cytology using a 22-gauge needle under ultrasound guidance. Light chain restriction was defined as a κ to λ ratio of less than 0.5 or more than 3.0. According to the light chain-positive rate, 25% or less and more than 25% were classified as the low and high light chain-positive rate groups, respectively. B-cell predominance was defined as a CD19 to CD4 ratio (B- to T-cell ratio) of more than 2.0. B-cell predominance was more frequently observed in lymphomas (88.5%) than in benign lymphoproliferative lesions (25.0%; p < 0.001). Light chain restriction based on the κ/λ ratio was detected in 69.2% of lymphomas, but not in benign lymphoproliferative lesions. Among lymphomas belonging to the low light chain-positive rate group, 88.9% did not exhibit light chain restriction and B-cell predominance was present. In contrast, benign lymphoproliferative lesions with B-cell predominance were not detected in the low light chain-positive rate group. B-cell predominance was a useful indicator for diagnosing thyroid lymphoma in the low light chain-positive rate group without light chain restriction.

Utility of a corticotropin-releasing hormone test to differentiate pituitary-dependent hyperadrenocorticism from cortisol-producing adrenal tumors in dogs.

BACKGROUND: Hyperadrenocorticism (HAC) is a common endocrine disorder in dogs; however, there are no reports on the use of the corticotropin-releasing hormone test (CRHT) to differentiate between pituitary-dependent hyperadrenocorticism (PDH) and cortisol-producing adrenal tumors (CPATs), both causative of HAC. OBJECTIVES: To evaluate the usefulness of CRHT as a tool to differentiate between PDH and CPAT in dogs and to determine the reference intervals for CRHT in healthy, PDH, and CPAT dogs. ANIMALS: Dogs diagnosed with PDH (n = 21), CPAT (n = 6), and healthy beagle dogs (n = 33). METHODS: This prospective study included dogs with a definitive diagnosis of PDH and CPAT and healthy beagle dogs, in which CRHT was performed, were prospectively evaluated. We investigated the correlations of CRHT (endogenous adrenocorticotropic hormone [ACTH] concentration, endogenous ACTH concentration [EAC], and poststimulation ACTH concentration [PAC]) with pituitary-to-brain ratio (PBR) (in PDH) and with indices of adrenal ultrasonography (smaller and larger adrenal gland dorsoventral thickness in PDH and CPAT). RESULTS: For EAC, the area under the curve (AUC) was 0.95, with a cutoff value of 26.3 pg/mL (sensitivity: 90.62%, specificity: 87.50%). The AUC for PAC was 0.96 with a cutoff value of 54.5 pg/mL (sensitivity: 100.00%, specificity: 66.67%). The 95% reference interval for CRHT in healthy (control) dogs ranged 5.00 to 79.8 pg/mL (1.10-17.57 pmol/L) for EAC, and 1.92 to 153.42 pg/mL (0.42-33.78 pmol/L) for PAC. There was no significant correlation between PBR and CRHT, nor adrenal size and CRHT. CONCLUSIONS AND CLINICAL IMPORTANCE: CRHT appears to be a rapid and reliable test for differentiating PDH from CPAT in dogs.

Canine Leptospirosis Outbreak in Japan.

Canine leptospirosis was suspected in 11 dogs in Osaka Prefecture, Japan and 9 dogs died within a month, from October 12 to November 10, 2017. Eight of the dogs had been taken on walks along the same riverbed and 4 dogs lived in the same town. Logistic regression analysis between a comparative group and the incident cases group showed that the odds of leptospirosis infection was 13.3 times higher (p = 0.044) in the dogs taken on walks along the riverbed than in the dogs not being walked along the riverbed. It is suggesting that these walks had been a risk factor. Microscopic agglutination tests showed that antibody titers against Leptospira interrogans serovar Australis were 1:2,560 and 1:10,240 in 2 dogs. Therefore, L. interrogans serovar Australis was suspected to be the causative agent, for which no canine vaccine is available in Japan. These results suggested that L. interrogans serovar Australis can cause local outbreaks. The development of a canine vaccine against various serotypes might help reduce local infections. Leptospirosis is an important infectious disease of dogs and it is also a zoonotic disease.

Experimental water injection into lungs using an animal model: Verification of the diatom concentration test to diagnose drowning.

BACKGROUND: In forensic medicine, the diatom test is used to diagnose drowning. Drowning and postmortem immersion can be distinguished by calculating the ratio of diatom concentration in the lungs and drowning water (L/W ratio). However, this claim was based on the unproven hypothesis that diatoms may be concentrated in the lungs due to respiratory movements. This study was conducted to examine whether the L/W ratio increased with experimental water injection. METHODS: A total of two experiments was performed using 22 non-drowned cat carcasses found on dry land. First, for the experimental postmortem immersion, we soaked seven whole-body cat carcasses in pond water for an hour. Second, the pond water was experimentally injected one or ten times into each harvested lung from seven and eight cats, respectively. In the diatom test, two diatom species (Aulacoseira ambigua and Discostella asterocostata) that were dominantly observed in pond water as well as other diatom species were counted separately. The L/W ratios of each cat were calculated. Univariate linear regression analysis was performed to demonstrate the association among L/W ratios and the three categories of the experiments. The L/W ratios of the two experiments were compared with those of drowning or postmortem immersion cases of humans or cats. RESULTS: It was revealed that the clear L/W ratio differences between the three groups (experimental postmortem immersion<0.02, 1-injection < 0.2, 10-injection > 0.9 for all diatom counting) were with statistically significant as proven by the univariate regression analyses. In actual cases of cats and humans, L/W ratios were>0.4 for drowning and<0.04 for postmortem immersion. CONCLUSION: The L/W ratio increased with multiple experimental water injections into the lungs, thereby verifying the validity of the diatom concentration test to diagnose drowning. The diatom test can be used to distinguish between drowning and postmortem immersion in humans and cats by calculating the L/W ratio.

Clinical significance and cytological detection of tracheal puncture following thyroid fine-needle aspiration: A retrospective study.

BACKGROUND: There have been no detailed reports on tracheal puncture after thyroid fine-needle aspiration (FNA). This study aimed to discuss the cytological differential diagnoses of tracheal puncture after thyroid FNA and clarify its clinical significance. METHODS: Tracheal puncture was defined as aspiration of tracheal components, including ciliated cells, chondrocytes, and goblet cells. A history of air suction or cough during aspiration was also considered tracheal puncture. Among the 18,480 nodules from 13,813 patients that underwent thyroid FNA, 70 (0.38%) nodules with tracheal puncture were retrospectively examined. Eleven thyroglossal duct cysts (TGDCs) and seven bronchial cysts that could exhibit ciliated cells were included in the study to compare the cytological findings. RESULTS: Sixty-six (94.3%) nodules had no clinical evidence of complications during and after FNA. Of the nodules with tracheal puncture, 64.3%, 48.6%, and 51.4% nodules were <1.0 cm in size, located in the isthmus, and associated with calcification. Cytological examination showed that chondrocytes and ciliated cells were present in 94.3% and 32.9% nodules, respectively. Ciliated cells seen in nodules with tracheal puncture and TGDCs were non-degenerative, whereas those in bronchial cysts were degenerative. CONCLUSION: Tracheal puncture after thyroid FNA is rarely noticed clinically, does not cause serious conditions, and spontaneously resolves. This complication more likely occurs in small-calcified nodules in the isthmus. Chondrocytes are more reliable diagnostic clues than ciliated cells to indicate tracheal puncture cytologically.

Evaluation of E-Cadherin and ß-Catenin Immunoreactivity for Determining Undifferentiated Cells in Anaplastic Thyroid Carcinoma.

INTRODUCTION: An immunohistochemical study has occasionally been performed to diagnose anaplastic thyroid carcinoma (ATC). However, antibodies to confirm the undifferentiated nature of ATC have not yet been evaluated. The aim of this study was to evaluate E-cadherin and ß-catenin expressions in immunoreactivity to determine undifferentiated carcinoma cells in the diagnosis of ATC. METHODS: We immunohistochemically examined 29 ATCs, 30 poorly differentiated thyroid carcinomas (PDTCs), 22 well-differentiated thyroid carcinomas (WDTCs), and 3 squamous cell carcinomas. Antibodies for thyroid transcription factor-1 (TTF-1), paired-box gene 8 (PAX8), ß-catenin, and E-cadherin were used. RESULTS: All WDTCs tested positive for TTF-1, PAX8, and E-cadherin. The positive rates of TTF-1, PAX8, and E-cadherin were 93.3, 93.3, and 100%, respectively, in PDTCs and 17.2, 51.7, and 10.3%, respectively, in ATCs. WDTC expressed the lateral cell membrane staining for ß-catenin and E-cadherin, whereas PDTC showed circumferential cell membranous expression (fishnet pattern). ß-catenin cell membrane expression in ATCs is lost or discontinuous. Carcinoma cells with ß-catenin nuclear expression without cell membranous expression were scattered in 72.4% of ATCs but were not observed in the other carcinomas. CONCLUSION: We propose 3 immunohistochemical findings to determine undifferentiated carcinoma cells in the diagnosis of ATC: (1) ß-catenin nuclear expression with no or reduced cell membranous expression, (2) the loss or discontinuous pattern of E-cadherin expression, and (3) the loss of PAX8 nuclear expression.

Potential role of mobile rapid on-site evaluation® in thyroid fine-needle aspiration cytology to reduce delayed repeated aspiration.

Rapid on-site evaluation of fine-needle aspiration cytology is time-consuming and requires specialized cytopathology staff. Mobile Rose® is a newly developed device for rapid on-site evaluation of fine-needle aspiration cytology. This study aimed to investigate the potential role of Mobile Rose® in reducing delayed repeated aspiration of the thyroid. A total of 120 cytological samples were collected and observed using Mobile Rose® after fine-needle aspiration cytology between September and October 2020, with immediate assessment of minimal or no cell clusters after conventional smear preparation. After qualifying and scoring, needle washout materials were prepared using the BD CytoRichTM method and correlated with cytology results. The average turn-around time of Mobile Rose® was found to be 1.5 minutes. Sensitivity, specificity, positive predictive value, and negative predictive value were 94.4%, 100%, 100%, and 57.1%, respectively. False-negative results were attributed to small aggregates of cells that were difficult to distinguish from the background and artifacts. Mobile Rose® may represent an important innovation for rapid on-site evaluation that is fast, has high diagnostic performance, does not require the presence of specialized cytology staff, and can reduce delayed repeated aspiration of the thyroid gland. However, further minor improvements and confirmation are required.

Fine-needle aspiration of parathyroid adenomas: Indications as a diagnostic approach.

BACKGROUND: We aimed to determine the indication of fine-needle aspiration (FNA) for parathyroid adenoma (PA)-suspected nodules and the cytological features of PA, and to discuss the ancillary techniques for diagnostic confirmation. METHOD: Clinical, cytological, and histological examinations of 15 PA patients (4.0% of all PA resected patients) were conducted through FNA on 16 nodules. We also examined the cytological preparations of 10 follicular neoplasms (FNs) and 10 poorly differentiated thyroid carcinomas (PDTCs). RESULTS: FNA was performed to detect PA in nine (56.3%) nodules. The remaining seven (43.8%) nodules underwent FNA for lesions considered as thyroid nodules or lymph nodes. The levels of parathyroid hormone (PTH) in the aspiration needle washout fluid were observably high, except for that from one nodule with unsatisfactory FNA. Cytologically, the incidences of wedge pattern (86.7%) and salt and pepper chromatin (86.7%) in PAs were significantly higher than in FNs and PDTCs. In contrast, the appearance of colloid globules and nuclear grooves was less frequent than that of FNs and PDTCs. GATA-3 expression was intense in all PAs that immunocytochemistry were performed. Histologically, capsular invasion and/or laceration, tumor seeding, granulation tissue, and fibrosis were observed. CONCLUSIONS: When PA localization is unusual or inconclusive despite extensive imaging, FNA may be performed. We asserted that wedge pattern, salt and pepper chromatin, and the absence of colloid globules and nuclear grooves are diagnostic cytological indicators of PA rather than of FN or PDTC. We recommend PTH measurements using needle washout fluid for PA-suspected nodules, and immunocytochemistry with the GATA-3 antibody for cytologically PA-suspected nodules.