The relationship between corneal calcareous degeneration and various ophthalmic diseases in dogs: A retrospective study.

OBJECTIVE: To investigate ocular factors that influence the development of corneal calcareous degeneration (CCD) in dogs. ANIMALS AND PROCEDURES: The medical records of 96 eyes of dogs with CCD and 288 eyes without CCD were retrospectively reviewed. Dogs with evidence of causative systemic illness associated with CCD were excluded from the study. Logistic regression analysis was used to identify the ocular factors associated with the development of CCD. To identify the effect of phosphate-containing eyedrops on CCD, the application periods of phosphate-containing antiglaucoma eyedrops were compared between the glaucomatous eyes in the CCD and non-CCD groups. RESULTS: Increased age, brachycephalic breed, keratoconjunctivitis sicca, advanced cataract, history of phacoemulsification, and topical corticosteroid application were significantly associated with CCD development. Glaucoma was significantly overrepresented in the non-CCD group, and the application period of phosphate-containing antiglaucoma eyedrops was significantly longer in eyes with CCD than in those without CCD. CONCLUSIONS: Ophthalmic diseases requiring long-term management of ocular inflammation and long-term application of phosphate-containing eyedrops may contribute to the development of CCD. Glaucoma is overrepresented in dogs without CCD, which is thought to be due to the differences in predisposed age and breeds between dogs with glaucoma and CCD.

Comparison of the outcomes of phacoemulsification versus topical medication alone in canine diabetic cataracts: a retrospective study.

BACKGROUND: Long-term comparisons of phacoemulsification with topical medication are limited in canine diabetic cataracts. OBJECTIVES: To compare outcomes of eyes submitted to phacoemulsification with those of topical medication for canine diabetic cataracts and identify risk factors for complications. METHODS: Through medical records review, 150 eyes (76 dogs) with diabetic cataracts were included; 58 eyes (31 dogs) underwent phacoemulsification (phaco-group) and 92 eyes (48 dogs) received ophthalmic solution alone (medication-group). The medication-group was divided into owner-led and vet-led groups depending on who elected not to perform surgery. Comparisons involved time-to-complications, vision, and the number and type of ophthalmic solutions administered. The association between complications and pretreatment clinical findings was investigated. RESULTS: No difference was found in complication risk between the phaco and owner-led medication groups. Conversely, the vet-led medication-group had a higher complication risk than the other groups. At the last follow-up, 94.8% of the phaco-group had vision, whereas 7.6% of the medication-group restored some visual axis. Poor glycemic control in the medication-group and younger age in the phaco-group increased complication risk. At 1-year post-treatment, the average number of ophthalmic solutions administered was 1.7 and 2.6 in the phaco and medication groups, respectively. The medication-group used anti-inflammatories the most throughout the follow-up, whereas the phaco-group used anti-inflammatories the most until 1-year post-treatment and lacrimostimulants at 1.5-year post-treatment. CONCLUSIONS: For canine diabetic cataracts, phacoemulsification is recommended because it is superior to topical management alone in terms of maintaining vision and reducing the number of ophthalmic solutions required in the long term.

Hema-seq reveals genomic aberrations in a rare simultaneous occurrence of hematological malignancies.

Co-occurrence of multiple myeloma and acute myelogenous leukemia is rare, with both malignancies often tracing back to multipotent hematopoietic stem cells. Cytogenetic techniques are the established baseline for diagnosis and characterization of complex hematological malignancies. In this study, we develop a workflow called Hema-seq to delineate clonal changes across various hematopoietic lineages through the integration of whole-genome sequencing, copy-number variations, cell morphology, and cytogenetic aberrations. In Hema-seq, cells are selected from Wright-stained slides and fluorescent probe-stained slides for sequencing. This technique therefore enables direct linking of whole-genome sequences to cytogenetic profiles. Through this method, we mapped sequential clonal alterations within the hematopoietic lineage, identifying critical shifts leading to myeloma and acute myeloid leukemia (AML) cell formations. By synthesizing data from each cell lineage, we provided insights into the hematopoietic tree's clonal evolution. Overall, this study highlights Hema-seq's capability in deciphering genomic heterogeneity in complex hematological malignancies, which can enable better diagnosis and treatment strategies.

Diagnosis of traumatic scleral perforation in a dog using ocular ultrasound and ultrasound biomicroscopy.

A 10-year-old spayed female Maltese dog was presented for hypotony of the right eye (OD) with decreased vision. The dorsomedial region of conjunctiva was partially swollen but the globe remained intact. Slit-lamp biomicroscopy revealed intraocular haemorrhage, uveitis, and a limbal, corneoconjunctival ulcer near the swollen conjunctiva. During ocular ultrasonography (OUS), the sclera became indented and hypotony occurred, implicating a full-thickness scleral wound. Ultrasound biomicroscopy (UBM) confirmed a full-thickness scleral defect, which was surgically apposed. The intraocular pressure normalised and the uveitis subsided post-operatively. OUS and UBM were helpful in suggesting and definitively diagnosing scleral perforation, leading to accurate treatment and good prognosis.

Distinct mutational pattern of T-cell large granular lymphocyte leukemia combined with pure red cell aplasia: low mutational burden of STAT3.

T-cell large granular lymphocyte leukemia (T-LGL) is often accompanied by pure red cell aplasia (PRCA). A high depth of next generation sequencing (NGS) was used for detection of the mutational profiles in T-LGL alone (n = 25) and T-LGL combined with PRCA (n = 16). Beside STAT3 mutation (41.5%), the frequently mutated genes included KMT2D (17.1%), TERT (12.2%), SUZ12 (9.8%), BCOR (7.3%), DNMT3A (7.3%), and RUNX1 (7.3%). Mutations of the TERT promoter showed a good response to treatment. 3 of 41 (7.3%) T-LGL patients with diverse gene mutations were revealed as T-LGL combined with myelodysplastic syndrome (MDS) after review of bone marrow slide. T-LGL combined with PRCA showed unique features (low VAF level of STAT3 mutation, low lymphocyte count, old age). Low ANC was detected in a STAT3 mutant with a low level of VAF, suggesting that even the low mutational burden of STAT3 is sufficient for reduction of ANC. In retrospective analysis of 591 patients without T-LGL, one MDS patient with STAT3 mutation was revealed to have subclinical T-LGL. T-LGL combined with PRCA may be classified as unique subtype of T-LGL. High depth NGS can enable sensitive detection of concomitant MDS in T-LGL. Mutation of the TERT promoter may indicate good response to treatment of T-LGL, thus, its addition to an NGS panel may be recommended.

Two different procedures for managing ocular melanosis presenting scleral pigmentation with glaucoma in a Shih-Tzu dog: A case of definitive diagnosis in one eye and a presumed diagnosis in the contralateral eye.

A 12-year-old castrated male Shih-Tzu dog was referred for uncontrolled glaucoma and uveitis with highly pigmented sclera, in both eyes (OU). On ophthalmic examination, the menace response, dazzle reflex and pupillary light reflex were negative OU. The intraocular pressure was 27 mmHg in the right eye (OD) and 70 mmHg in the left eye (OS) despite the administration of antiglaucoma eyedrops. Ultrasound biomicroscopy revealed a closed ciliary cleft OU. Ocular ultrasonography revealed hyperechoic materials in the vitreous OU and retinal detachment OS. When presented for recheck, an extensive malacic corneal ulcer was observed OS. To relieve pain in the blind eyes, enucleation OS and pharmacologic ciliary body ablation (CBA) OD were performed. Histologically, ocular melanosis, which is an inherited disease in the Cairn Terrier breed, was identified in the enucleated eye. The uvea was heavily pigmented. The iris and ciliary body were mildly distorted by a single population of large, round, nonneoplastic cells with pigmented cytoplasm. There was no evidence of an intraocular mass or metastasis before and after intravitreal CBA. This is the first report of bilateral ocular melanosis in a Shih-Tzu dog. Ocular melanosis is a possible differential diagnosis for globe presenting scleral pigmentation with glaucoma in even non-Cairn Terrier breeds and pharmacologic CBA could be considered as a treatment for ocular melanosis with end-stage glaucoma.

Analysis of clinical and genomic profiles of therapy-related myeloid neoplasm in Korea.

BACKGROUND: Therapy-related myeloid neoplasm (T-MN) rarely occurs among cancer survivors, and was characterized by poor prognosis. T-MN has germline predisposition in a considerable proportion. Here, clinical characteristics and germline/somatic variant profiles in T-MN patients were investigated, and the findings were compared with those of previous studies. METHODS: A review of medical records, cytogenetic study, targeted sequencing by next-generation sequencing, and survival analysis were performed on 53 patients with T-MN at a single institution in Korea. RESULTS: The patients were relatively younger compared to T-MN patients in other studies. Our T-MN patients showed a high frequency of complex karyotypes, -5/del(5q), and -7/del(7q), which was similar to the Japanese study group but higher than the Australian study group. The most common primary disease was non-Hodgkin lymphoma, followed by breast cancer. The detailed distributions of primary diseases were different across study groups. Seven patients (13.2%) harbored deleterious presumed/potential germline variants in cancer predisposition genes (CPG) such as BRIP1, CEBPA, DDX41, FANCM, NBN, NF1, and RUNX1. In the somatic variant profile, TP53 was the most frequently mutated gene, which was consistent with the previous studies about T-MN. However, the somatic variant frequency in our study group was lower than in other studies. Adverse factors for overall survival were male sex, older age, history of previous radiotherapy, previous longer cytotoxic therapy, and -5/del(5q). CONCLUSION: The findings of our study corroborate important information about T-MN patients. As well as a considerable predisposition to CPG, the clinical characteristics and somatic variant profile showed distinctive patterns. Germline variant testing should be recommended for T-MN patients. If the T-MN patients harbor pathogenic germline variants, the family members for stem cell donation should be screened for carrier status through germline variant testing to avoid donor-derived myeloid neoplasm. For the prediction of the prognosis in T-MN patients, sex, age, past treatment history, and cytogenetic findings can be considered.

Dual regulation of Kv7.2/7.3 channels by long-chain n-alcohols.

Normal alcohols (n-alcohols) can induce anesthetic effects by acting on neuronal ion channels. Recent studies have revealed the effects of n-alcohols on various ion channels; however, the underlying molecular mechanisms remain unclear. Here, we provide evidence that long-chain n-alcohols have dual effects on Kv7.2/7.3 channels, resulting in channel activation as the net effect. Using heterologous expression systems, we found that n-alcohols could differentially regulate the Kv7.2/7.3 channel depending on their chain length. Treatment with short-chain ethanol and propanol diminished Kv7.2/7.3 currents, whereas treatment with long-chain hexanol and octanol enhanced the currents. However, the long-chain alcohols failed to potentiate Kv7.2 currents pre-activated by retigabine. Instead, they inhibited the currents, similar to short-chain ethanol. The stimulatory effect of the long-chain n-alcohols was also converted into an inhibitory one in the mutant Kv7.2(W236L) channels, while the inhibitory effect of ethanol did not differ between wild-type Kv7.2 and mutant Kv7.2(W236L). The inhibition of currents by n-alcohols was also seen in Kv7.1 channel which does not have the tryptophan (W) residue in S5. These findings suggest that long-chain n-alcohols exhibit dual effects through independent working sites on the Kv7.2 channel. Finally, we confirmed that the hydroxyl group with a negative electrostatic potential surface is essential for the dual actions of n-alcohol. Together, our data suggest that long-chain n-alcohols regulate Kv7.2/7.3 channels by interacting with both stimulatory and inhibitory sites and that their stimulatory action depends on the conserved tryptophan 236 residue in S5 and could be important for triggering their anesthetic effects.

Novel THPO variant in hereditary thrombocytopenia: A potential candidate variant for predisposition to myeloid neoplasm.

Hereditary thrombocytopenia is a heterogeneous group of congenital disorders with a wide range of symptoms depending on the severity of platelet dysfunction or thrombocytopenia. Because of its clinical phenotypes and the bone marrow morphology associated with this condition, hereditary thrombocytopenia can be misdiagnosed as primary immune thrombocytopenia and myelodysplastic syndrome. Therefore, genetic evidence is necessary for the accurate diagnosis of hereditary thrombocytopenia. Refractory cytopenia of childhood is a subgroup of myelodysplastic syndrome that was added to the World Health Organization classification in 2008. To investigate the germline and somatic variants associated with refractory cytopenia of childhood, we performed targeted multigene sequencing in three patients with refractory cytopenia of childhood. Of the three patients, one progressed from megakaryocytic hypoplasia with thrombocytopenia, and targeted multigene sequencing revealed THPO variants in this patient and his sister. We propose that the monoallelic deletion of THPO is a potential candidate for germline predisposition to myeloid malignancy.

Assessment of Rotational Thromboelastometry and Thrombin Generation Assay to Identify Risk of High Blood Loss and Re-Operation After Cardiac Surgery.

Introduction: We aimed to investigate parameters for prediction of post-operative blood loss and re-operation in patients who underwent cardiopulmonary bypass. Methods: Thrombin generation assay, activated partial thromboplastin time, activated clotting time and rotational thromboelastometry (ROTEM) tests were performed at 4 time points in 65 patients: before skin incision (T1), after heparin injection (T2), after protamine reversal (T3) and before skin closure (T4). Results: Pre-operative endogenous thrombin potential (ETP) and peak thrombin levels were significantly lower in patients with high post-operative blood loss (≥ 800 mL) within 24 h than in those with low blood loss (< 800 mL). Clotting time (CT), maximal clotting firmness, clotting firmness time and alpha angle values of ROTEM measured at T2, T3 or T4 were significant predictors for high post-operative blood loss. An increase in CT-EXTEM over 4 time points was significant in patients who had a re-operation within 48 h compared to their counterparts. Conclusions: This study indicates that pre-operative ETP could predict high post-operative blood loss and that intra-operative ROTEM also helps to stratify risks of high post-operative blood loss and re-operation.

Heterogeneous genetic landscape of congenital neutropenia in Korean patients revealed by whole exome sequencing: genetic, phenotypic and histologic correlations.

Congenital neutropenia (CN) is a hematological disease heterogeneous in its genetic, phenotypic and histologic aspects. We aimed to identify the genetic etiology of Korean CN patients in the context of bone marrow (BM) histology and clinical phenotype. Whole-exome sequencing (WES) or targeted sequencing was performed on the BM or peripheral blood specimens of 16 patients diagnosed with CN based on BM exam from 2009 to 2018. Absolute count of myeloperoxidase (MPO)-positive cells was calculated using ImageJ software. Semi-quantitation of MPO-positive cells in BM sections was performed by MPO grading (grades 0-3). Comprehensive retrospective review on real-world data of 345 pediatric patients with neutropenia including 16 patients in this study during the same period was performed. Seven disease-causing variants were identified in ELANE, G6PC3 and CXCR4 in 7 patients. A novel homozygous G6PC3 variant (K72fs) of which the mechanism was copy-neutral loss of heterozygosity was detected in two brothers. A low myeloid-to-erythroid ratio (0.5-1.5) was consistently observed in patients with ELANE mutations, while MPO-positive cells (40%-50%) with MPO grade 1 or 2 were detected in myelokathexis caused by G6PC3 and CXCR4 mutations. Meanwhile, disease-causing variants were detected in ELANE, TAZ and SLC37A4 in 5 patients by retrospective review of medical records. Our results suggest that following the immunological study and BM exam, WES or an expanded next generation sequencing panel that covers genes related to immunodeficiency and other inherited bone marrow failures as well as CN is recommended for neutropenia patient diagnosis.

Clinical and Genomic Profiles of Korean Patients with MECOM Rearrangement and the t(3;21)(q26.2;q22.1) Translocation.

The translocation (3;21)(q26.2;q22.1) is a unique cytogenetic aberration that characterizes acute myeloid leukemia with myelodysplasia-related changes (AML-MRC) in patients with AML and myelodysplastic syndrome (MDS) or a therapy-related myeloid neoplasm. Using multigene target sequencing and FISH, we investigated the clinical and genomic profiles of patients with t(3;21) over the past 10 years. The frequency of t(3;21) among myeloid malignancies was very low (0.2%). Half of the patients had a history of cancer treatment and the remaining patients had de novo MDS. Twenty-one somatic variants were detected in patients with t(3;21), including in CBL, GATA2, and SF3B1. Recurrent variants in RUNX1 (c.1184A>C, p.Glu395Ala) at the same site were detected in two patients. None of the patients with t(3;21) harbored germline predisposition mutations for myeloid neoplasms. MECOM rearrangement was detected at a higher rate using FISH than using G-banding, suggesting that FISH is preferable for monitoring. Although survival of patients with t(3;21) is reportedly poor, the survival of patients with t(3;21) in this study was not poor when compared with that of other AML patients in Korea.

Statistical Seasonal Forecasting of Winter and Spring PM2.5 Concentrations Over the Korean Peninsula.

Concentrations of fine particulate matter smaller than 2.5 µm in diameter (PM2.5) over the Korean Peninsula experience year-to-year variations due to interannual variation in climate conditions. This study develops a multiple linear regression model based on slowly varying boundary conditions to predict winter and spring PM2.5 concentrations at 1-3-month lead times. Nation-wide observations of Korea, which began in 2015, is extended back to 2005 using the local Seoul government's observations, constructing a long-term dataset covering the 2005-2019 period. Using the forward selection stepwise regression approach, we identify sea surface temperature (SST), soil moisture, and 2-m air temperature as predictors for the model, while rejecting sea ice concentration and snow depth due to weak correlations with seasonal PM2.5 concentrations. For the wintertime (December-January-February, DJF), the model based on SSTs over the equatorial Atlantic and soil moisture over the eastern Europe along with the linear PM2.5 concentration trend generates a 3-month forecasts that shows a 0.69 correlation with observations. For the springtime (March-April-May, MAM), the accuracy of the model using SSTs over North Pacific and 2-m air temperature over East Asia increases to 0.75. Additionally, we find a linear relationship between the seasonal mean PM2.5 concentration and an extreme metric, i.e., seasonal number of high PM2.5 concentration days. Supplementary Information: The online version contains supplementary material available at 10.1007/s13143-022-00275-4.

Evaluation of ocular surface parameters in dogs with and without meibomian gland dysfunction.

BACKGROUND: Interest in meibomian gland dysfunction (MGD) is growing in veterinary medicine. However, research on MGD in dogs is lacking. The aims of this study were to compare the interferometry grades, tear meniscus height (TMH) and non-invasive tear break-up time (NIBUT) grades between dogs with and without MGD. METHODS: Routine ophthalmic examination, interferometry, NIBUT assessment, TMH measurement and meibography were performed. Age and Schirmer tear test-1 (STT-1) results were compared between the control and MGD groups using Student's t-test. Interferometry grades, NIBUT grades and TMH were compared between the two groups using Mann-Whitney U-test. RESULTS: There was no significant difference in age between the two groups (p = 0.279). STT-1 (p = 0.024), interferometry (p = 0.004) and NIBUT grades (p = 0.012) were significantly lower in the MGD group than in the control group. No significant difference in TMH values (p = 0.587) was observed between the two groups. While the control group included 18 and seven eyes in meiboscore 0 and 1, in MGD group, 12, eight, five and three eyes were included in meiboscore 0, 1, 2 and 3, respectively. CONCLUSIONS: Low interferometry and NIBUT grades were associated with MGD, suggesting decreased meibum and disrupted tear film quality. TMH did not differ between the two groups. Meibography could aid in the diagnosis of MGD in severe cases, although it could not detect early MGD in the dogs in this study.

Quantification of tear glucose levels and their correlation with blood glucose levels in dogs.

BACKGROUND: No previous studies have quantified tear glucose (TG) levels in dogs or compared changes in TG and blood glucose (BG) concentrations. OBJECTIVE: To quantify TG concentration and evaluate its correlation with BG level in dogs. METHODS: Twenty repetitive tests were performed in alternate eyes of four dogs, with a minimum washout period of 1 week. Tears and blood were collected at 30-min intervals with successive glucose injections (1 g/kg) every 30 min. Cross-correlations of BG and TG levels were assessed. The delay and association between TG and corresponding BG levels were analysed for each dog; samples were collected at 5-min intervals. The tears were collected using microcapillary tubes. Collected tears and blood were analysed for glucose concentration using a colorimetric assay and commercially available glucometer, respectively. RESULTS: The average baseline BG and TG levels were 4.76 ± 0.58 and 0.39 ± 0.04 mmol/L, respectively. Even with highly fluctuating BG levels, a significant cross-correlation coefficient (r = 0.86, p < 0.05) was observed between changes of BG and TG levels. The delay time between BG and TG levels was 10 min. On average, BG levels were 16.34 times higher than TG levels. There was strong correlation between BG and TG levels (rs  = 0.80, p < 0.01). Significant differences in TG concentrations between normoglycaemia, mild hyperglycaemia, and severe hyperglycaemia were found (p < 0.05). CONCLUSIONS: Canine TG concentrations have not been quantified previously. Our findings suggest preliminary data for future research on TG levels in dogs and show TG measurement could be used to screen for diabetes mellitus in dogs.

Genetic profile of primary plasma cell leukemia in Korea: comparison with plasma cell myeloma.

Plasma cell leukemia (PCL) is clinically and genetically distinct from multiple myeloma (MM), despite controversies regarding the disease definition. To determine the distinct features of PCL, the genetic property of primary PCL (pPCL) was compared with that of secondary PCL (sPCL) and MM. In patients with pPCL, Eighty-nine non-synonymous mutations were observed in 68 genes. The most frequently mutated genes were TP53, TSC2, and TYK2. In comparison with genetic abnormalities of sPCL and MM, 45 genes were present only in pPCL while 28 genes were only in sPCL and 22 genes only in MM. Among the common genes between pPCL and MM, a higher prevalence of TP53 was observed in pPCL, compared to MM (p < 0.05), while similar, compared to sPCL (p = 0.64). In summary, pPCL patients showed a higher level of genetic heterogeneity and distinctive genetic signature in their mutational profile compared to patients with MM and sPCL.

Comparison of iridocorneal angle parameters measured by spectral domain optical coherence tomography and ultrasound biomicroscopy in dogs.

OBJECTIVE: To compare the measurements of iridocorneal angle parameters between spectral domain optical coherence tomography (SD-OCT) and ultrasound biomicroscopy (UBM) in dogs. ANIMALS STUDIED AND PROCEDURES: A total of 47 eyes of dogs were scanned at the temporal limbus using SD-OCT and UBM. Iridocorneal angle (ICA) and angle opening distance (AOD) were measured from the obtained images accordingly. The intra-observer and inter-observer reproducibility were evaluated using the intraclass correlation coefficient. To evaluate intra-observer reproducibility, measurements of the first and second grading from the first examiner were compared. To evaluate inter-observer reproducibility, measurements between the two examiners were compared. Agreement between ICA and AOD for SD-OCT and UBM was evaluated using Bland-Altman plots. RESULTS: In the first grading, the mean ICA and AOD for SD-OCT were 31.4 ± 6.4° and 641.4 ± 270.8 µm, respectively. The mean ICA and AOD for UBM were 32.0 ± 4.8° and 700.4 ± 238.8 µm, respectively. For ICA and AOD measurements, intra-observer reproducibility was excellent for both devices, whereas inter-observer reproducibility was excellent for SD-OCT and good for UBM. The mean difference in ICA between SD-OCT and UBM was 0.6° with a limit of agreement (LoA) span of 18.9°. The mean difference in AOD between SD-OCT and UBM was 58.9 µm with a LoA span of 804.4 µm. CONCLUSIONS: Spectral domain optical coherence tomography is an effective non-contact imaging modality for the evaluation of canine iridocorneal angle parameters in a clinical setting. Reproducibility of measurements obtained is comparable or superior to UBM, but values obtained by SD-OCT and UBM for AOD are not interchangeable between devices.