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We report a case of Becker muscular dystrophy in a 6-month-old, mixed-breed, castrated male pig detected with macroglossia at a meat inspection center. The pig presented a severely enlarged tongue extending outside its mouth. The tongue was firm and pale with discolored muscles. Histologically, there was severe fibrosis, fatty replacement, and myofiber necrosis, degeneration, and regeneration. Immunofluorescence showed focal and severely weak labeling for dystrophin at the sarcolemma of myocytes in the tongue. Analysis of dystrophin mRNA showed a 62 base pair insertion between exons 26 and 27. The insertion was derived from intron 26. Based on these findings, we diagnosed the case as Becker muscular dystrophy—the first known muscular dystrophy case induced by pseudoexon insertion in animals.

There is no nationwide necropsy database of animals in Japan, and most of the records are available from the postwar period. To clarify the chronological transition of animal necropsy cases in Tokyo, Japan, the records accumulated in the University of Tokyo from 1902 were investigated. Of necropsy records on paper or electronic from 1902 to 2021 kept at the Laboratory of Veterinary Pathology, the University of Tokyo, totally 3137 cases including 572 in 1903-1914 (the Meiji-Taisho period), 1258 in 1956-1969 (the Showa period) and 1307 in 2006-2020 (the Heisei-Reiwa period), respectively, were examined for species, breed, age and pathological diagnosis. Dogs (44.6%) and horses (34.8%) in the Meiji-Taisho period, dogs (62.9%) and cats (17.3%) in the Showa period, and dogs (46.0%), cats (26.1%) and exotic animals (20.5%) in the Heisei-Reiwa period were the most necropsied animal species. With the passage of time, the number of animal species increased, and the breeds of dogs and cats came to be more various. The median ages of death were 2 years, 3 years and 10 years old in dogs in the Meiji-Taisho, Showa and Heisei-Reiwa periods, respectively, and 2 years and 10 years old in cats in the Showa and Heisei-Reiwa periods, respectively. Viral, bacterial and parasitic infections were decreased, and inversely tumor cases increased due to the prolonged lifespan.

Aims Our objective was to develop a population pharmacokinetic (PK) model to describe roxadustat plasma concentrations in Japanese dialysis-dependent chronic kidney disease (DD-CKD) patients with renal anemia and to identify the covariate factors that affect exposure of roxadustat. Methods Three hundred sixty-seven patients (male, 256; female, 111) contributing 1285 concentration values from four clinical studies were analyzed using a nonlinear mixed-effects modeling approach. Candidate covariates included clinical characteristics hypothesized to affect roxadustat clearance and bioavailability, such as demographics, hepatic parameters, and concomitant drugs. Results The roxadustat PK data in Japanese DD-CKD patients with renal anemia were well described by a two-compartment disposition model with first-order absorption and interindividual variability on clearance, central volume of distribution, and absorption rate constant. Age was identified as a significant covariate on clearance. PK profiles of hemodialysis and peritoneal dialysis patients were comparable. Eighty-two percent of patients were administered at least one phosphate binder (PB). The effect of PBs on roxadustat concentration was modeled as a decrease in bioavailability. Staggered administration of PBs reduced the effect on roxadustat bioavailability. The clinical impact of all covariates on roxadustat PK was mild and manageable as the roxadustat dose was titrated based on hemoglobin level and administered starting from a low dose. Conclusion Roxadustat PK in Japanese DD-CKD patients were successfully described by a population PK model. The identified key covariates included coadministration of PBs on the roxadustat bioavailability and age on clearance of roxadustat.

Background: Rabbits, ferrets, and four-toed hedgehogs are popular exotic pets, but comprehensive epidemiological studies in these animals have been rarely conducted. The present study aims to clarify the incidence of neoplastic and non-neoplastic diseases in pet rabbits, ferrets and four-toed hedgehogs in Japan. Methods: We histologically/cytologically investigated 1098 samples from 883 rabbits, 812 samples from 521 ferrets and 561 samples from 468 four-toed hedgehogs that were collected at Miwa Exotic Animal Hospital and submitted to the Laboratory of Veterinary Pathology, The University of Tokyo between 2006 and 2020. The examinations of necropsy samples were not included in the present study. Results: Of the 1098 samples from diseased rabbits, 721 (65.7%) were diagnosed as neoplastic and 377 (34.3%) were as non-neoplastic. Uterine adenocarcinoma (21.1%), cutaneous soft tissue sarcoma (15.1%) and mammary gland adenocarcinoma (8.2%) were the most commonly encountered neoplasms. Endometrial hyperplasia (35.8%), testicular atrophy (4.2%) and uterine adenomyosis (3.7%) were the most common non-neoplastic lesions in rabbits. Of the 812 samples from diseased ferrets, 487 (60.0%) were diagnosed as neoplastic and 325 (40.0%) were as non-neoplastic. Adrenocortical tumor (23.2%), lymphoma (19.3%) and pancreatic islet cell tumor (11.5%) were the most commonly encountered neoplasms. Extramedullary hematopoiesis in the spleen (10.5%), lymph node hyperplasia (7.7%) and cholangiohepatitis (4.6%) were the most common non-neoplastic lesions in ferrets. Of the 561 samples from diseased hedgehogs, 338 (60.2%) were diagnosed as neoplastic and 223 (39.8%) were as non-neoplastic. Endometrial stromal tumor and endometrial mixed tumor (23.7%), oral squamous cell carcinoma (13.0%) and cutaneous soft tissue sarcoma (11.5%) were the most commonly encountered neoplasms. Gingivitis/stomatitis (39.5%), endometrial hyperplasia (18.8%) and dermatitis (6.7%) were the most common non-neoplastic lesions in hedgehogs. Conclusions and clinical relevance: Information obtained from the present study will provide a useful reference for veterinarians working with these exotic animals. Based on the literature search, this is the largest-scale retrospective study on disease incidence in hedgehogs.

Mammalian orthoreovirus (MRV) is a non-enveloped double-stranded RNA virus with a broad host range. MRVs are prevalent worldwide and have been isolated from various hosts, including humans, dogs, cats, wild boars, sewage, and pigs, in Japan. However, Japanese porcine MRV has not been genetically characterized yet. While investigating porcine enteric viruses including MRV, five MRVs were isolated from the feces of Japanese pigs using the MA104 cell culture. Genetic analysis of the S1 gene revealed that the Japanese porcine MRV strain isolates could be classified as MRV-2 and MRV-3. Whole genome analysis showed that Japanese porcine MRVs exhibited genetic diversity, although they shared homology with porcine MRV sequences deposited in the DDBJ/EMBL/GenBank database. Several potential intra-genetic reassortment events among MRV strains from pigs, sewage, and humans have been detected in Japan, suggesting zoonotic transmission. Furthermore, homologous recombination events were identified in the M1 and S1 genes of Japanese porcine MRV. These findings imply that different strains of Japanese porcine MRVs share a porcine MRV genomic backbone and have evolved through intra-genetic reassortment and homologous recombination events.

Rhabdomyosarcoma (RMS) is an aggressive type of soft tissue sarcoma, and pleomorphic RMS is a rare subtype of RMS found in adult. p16 is a tumor suppressor which inhibits cell cycle. In human RMS, p16 gene is frequently deleted, but p16-null mice do not develop RMS. We reported that genetic ablation of p16 by the crossbreeding of p16 knock-out rats (p16-KO rats) improved the dystrophic phenotype of a rat model of Duchenne muscular dystrophy (Dmd-KO rats). However, p16/Dmd double knock-out rats (dKO rats) unexpectedly developed sarcoma. In the present study, we raised p16-KO, Dmd-KO, and dKO rats until 11 months of age. Twelve out of 22 dKO rats developed pleomorphic RMS after 9 months of age, while none of p16-KO rats and Dmd-KO rats developed tumor. The neoplasms were connected to skeletal muscle tissue with indistinct borders and characterized by diffuse proliferation of pleomorphic cells which had eosinophilic cytoplasm and atypical nuclei with anisokaryosis. For almost all cases, the tumor cells immunohistochemically expressed myogenic markers including desmin, MyoD, and myogenin. The single cell cloning from tumor primary cells gained 20 individual Pax7-negative MyoD-positive RMS cell clones. Our results demonstrated that double knock-out of p16 and dystrophin in rats leads to the development of pleomorphic RMS, providing an animal model that may be useful to study the developmental mechanism of pleomorphic RMS.

Duchenne muscular dystrophy (DMD) is X-linked recessive myopathy caused by mutations in the dystrophin gene. Although conventional treatments have improved their prognosis, inevitable progressive cardiomyopathy is still the leading cause of death in patients with DMD. To explore novel therapeutic options, a suitable animal model with heart involvement has been warranted.We have generated a rat model with an out-of-frame mutation in the dystrophin gene using CRISPR/Cas9 genome editing (DMD rats). The aim of this study was to evaluate their cardiac functions and pathologies to provide baseline data for future experiments developing treatment options for DMD.In comparison with age-matched wild rats, 6-month-old DMD rats showed no significant differences by echocardiographic evaluations. However, 10-month-old DMD rats showed significant deterioration in left ventricular (LV) fractional shortening (P = 0.024), and in tissue Doppler peak systolic velocity (Sa) at the LV lateral wall (P = 0.041) as well as at the right ventricular (RV) free-wall (P = 0.004). These functional findings were consistent with the fibrotic distributions by histological analysis.Although the cardiac phenotype was milder than anticipated, DMD rats showed similar distributions and progression of heart involvement to those of patients with DMD. This animal may be a useful model with which to develop effective drugs and to understand the underlying mechanisms of progressive heart failure in patients with DMD.

Duchenne muscular dystrophy (DMD) is a progressive disease characterised by chronic muscle degeneration and inflammation. Our previously established DMD model rats (DMD rats) have a more severe disease phenotype than the broadly used mouse model. We aimed to investigate the role of senescence in DMD using DMD rats and patients. Senescence was induced in satellite cells and mesenchymal progenitor cells, owing to the increased expression of CDKN2A, p16- and p19-encoding gene. Genetic ablation of p16 in DMD rats dramatically restored body weight and muscle strength. Histological analysis showed a reduction of fibrotic and adipose tissues invading skeletal muscle, with increased muscle regeneration. Senolytic drug ABT263 prevented loss of body weight and muscle strength, and increased muscle regeneration in rats even at 8 months—the late stage of DMD. Moreover, senescence markers were highly expressed in the skeletal muscle of DMD patients. In situ hybridization of CDKN2A confirmed the expression of it in satellite cells and mesenchymal progenitor cells in patients with DMD. Collectively, these data provide new insights into the integral role of senescence in DMD progression.

Roxadustat is a hypoxia‐inducible factor prolyl hydroxylase inhibitor approved in China for anemia of dialysis‐dependent chronic kidney disease (CKD). Japanese hemodialysis patients with anemia of CKD previously naïve to, or converted from, erythropoiesis‐stimulating agents (ESAs) were enrolled in two open‐label, noncomparative studies of titrated oral roxadustat administered three times weekly. ESA‐naïve patients (n = 75) were randomized to roxadustat (initial dose, 50 or 70 mg) for 24 weeks; ESA‐converted patients (n = 164) were assigned to roxadustat (initial dose, 70 or 100 mg based on prior ESA dose) for 52 weeks. Efficacy outcomes included average hemoglobin (Hb, weeks 18‐24 or 46‐52), change of Hb from baseline to weeks 18 to 24 (ΔHb18‐24) or weeks 46 to 52 (ΔHb46‐52), and maintenance rate (proportion of patients who achieved average Hb of 10.0‐12.0 g/dL for weeks 18‐24 or weeks 46‐52). Treatment‐emergent adverse events (TEAEs) were monitored. Mean (SD) Hb was 10.93 (0.79) g/dL (weeks 18‐24) (ESA‐Naïve Study), and 10.93 (0.69; weeks 18‐24) g/dL and 11.11 (0.67; weeks 46‐52) g/dL (ESA‐Converted Study). Mean (SD) ΔHb18‐24 was 2.26 (1.02) g/dL (ESA‐Naïve Study) and −0.03 (0.90) g/dL (ESA‐Converted Study); mean (SD) ΔHb46‐52 was 0.12 (0.83) g/dL (ESA‐Converted Study). The overall maintenance rate was 73.0% (54/74) (ESA‐Naïve Study) (weeks 18‐24), and 79.1% (129/163; weeks 18‐24) and 71.2% (116/163; weeks 46‐52) (ESA‐Converted Study). Nasopharyngitis was the most common TEAE. Two deaths, considered unrelated to roxadustat, occurred in the ESA‐Converted Study. Roxadustat effectively corrected and maintained Hb, regardless of previous ESA treatment, in Japanese anemic CKD patients on hemodialysis.

The objective of this analysis was to develop and evaluate a pharmacokinetic/pharmacodynamic (PK/PD) model of the effect of roxadustat on low-density lipoprotein cholesterol (LDL-C) in Japanese patients with anemia of dialysis-dependent chronic kidney disease while considering the impact of covariates on model parameters.A total of 2330 LDL-C measurements from 275 patients in 3 clinical studies were analyzed using a nonlinear-mixed effects modeling approach in NONMEM software.The PK/PD relationship between roxadustat exposure and LDL-C was well described by a kinetic-pharmacodynamic model with a physiological indirect response model as the PD component. Co-administered statin usage, sevelamer usage, type of dialysis (hemodialysis or peritoneal dialysis), and sex were selected as covariates for LDLRoxadustat can decrease LDL-C independent of statins and sevelamer. Further study of the ability of roxadustat to lower LDL-C and any potential effects on outcomes is needed.

To clarify the prevalence of canine intracranial tumors in Japan, a retrospective study was performed using data on 186 canine intracranial tumors. Of 186 cases, 159 cases (85.5%) were primary and 27 cases (14.5%) were secondary intracranial tumors. Among primary intracranial tumors, meningioma (50.9%) was the most common, followed by glial tumors (21.4%) and primary intracranial histiocytic sarcoma (12.6%). These 3 tumors were most frequently found in middle-aged to elderly dogs without any sex predilection. Regarding glial tumors, the incidence of oligodendroglial tumors (79.4%) was higher than that of astrocytic tumors (17.6%). A significant breed predisposition (P

Duchenne muscular dystrophy (DMD) is a progressive muscular disorder caused by X-chromosomal DMD gene mutations. Recently, a new CRISPR/Cas9-mediated DMD rat model (cDMDR) was established and is expected to show cardiac lesions similar to those in humans. We therefore investigated the pathological and pathophysiological features of the cardiac lesions and their progression in cDMDR. For our cDMDR, Dmd-mutated rats (W-Dmd em1Kykn ) were obtained. Dmd heterozygous-deficient females and wild-type (WT) males were mated, and male offspring including WT as controls were used. (1) Hearts were collected at 3, 5, and 10 months of age, and HE- and Masson's trichrome-stained specimens were observed. (2) Electrocardiogram (ECG) recordings were made and analyzed at 3, 5, and 8 months of age. (3) Echocardiography was performed at 9 months of age. In cDMDR rats, (1) degeneration/necrosis of cardiomyocytes and myocardial fibrosis prominent in the right ventricular wall and the outer layer of the left ventricular wall were observed. Fibrosis became more prominent with aging. (2) Lower P wave amplitudes and greater R wave amplitudes were detected. PR intervals tended to be shorter. QT intervals were longer at 3 months but tended to be shorter at 8 months. Sinus irregularity and premature ventricular contraction were observed at 8 months. (3) Echocardiography indicated myocardial sclerosis and a tendency of systolic dysfunction. Pathological and pathophysiological changes occurred in cDMDR rat hearts and progressed with aging, which is, to some extent, similar to what occurs in humans. Thus, cDMDR could be a valuable model for studying cardiology of human DMD.

A 7‐year‐old neutered female domestic shorthaired cat born in Poland and then moved to Japan presented to the local clinic with recent onset of convulsive cluster seizures and status epilepticus. Magnetic resonance imaging revealed bilateral swelling of the hippocampus with T2 hyperintensity and contrast enhancing image, suggesting hippocampal necrosis. The cat completely recovered after treatment with antiepileptic drugs (AED) and administration of prednisolone (1 mg/kg PO q24h for 4 days and tapered). However, cluster seizures reoccurred and developed into status epilepticus despite increasing doses of AED. Although the convulsions were resolved by other AEDs, stupor and renal failure developed, and the cat was euthanized. Pathological findings were consistent with hippocampal necrosis. Immunological analysis for leucine‐rich glioma inactivated 1 (LGI1) autoantibodies was negative, but antibodies against DCC (deleted in colorectal carcinoma) known as netrin‐1 receptor were found. This report describes a case of feline autoimmune limbic encephalitis and hippocampal necrosis that were presumably associated with DCC autoantibodies.

Atypical porcine pestivirus (APPV), which has been confirmed to be associated with congenital tremor (CT) in pigs, is a newly discovered porcine virus that has been found in the Americas, Europe, and Asia; however, no report of APPV in Japan has been published. We identified an APPV in the central nervous system of Japanese piglets with CT, and firstly determined and analyzed the complete genome sequence. Phylogenetic analysis using the complete genome nucleotide sequence of the Japanese APPV, named Anna/2020, and those of APPVs from the NCBI database showed that APPVs were divided into three genotypes (genotypes 1 to 3), and that Anna/2020 clustered with the genotype 3 APPV strains, but distantly branched from these strains. Pairwise complete coding region nucleotide sequence comparisons revealed that there was 94.0% to 99.7% sequence identity among the genotype 3 strains, while Anna/2020 showed 87.0% to 89.3% identity to those genotype 3 strains, suggesting that Anna/2020 represents a novel APPV lineage within genotype 3. Retrospective examinations using RT-PCR revealed one genotype 1 and two novel genotype 3 APPVs from pigs without CT, and that novel genotype 3 APPVs have been prevalent in Japan since at least 2007.

A 6-year-old intact female sugar glider (Petaurus breviceps) had a swelling at the pouch region and died 8 days after presentation. At necropsy, the urinary bladder and abdominal lymph nodes were enlarged and a mass was located in the pericloacal region. Tumour infiltration was also observed in the medial iliac lymph nodes, liver, spleen, small intestine, uterus and left ovary. Histopathologically, the tumours were composed of pleomorphic histiocytes that had round or bizarre nuclei and abundant pale eosinophilic cytoplasm. Immunohistochemically, tumour cells were positive for ionized calcium-binding adaptor molecule 1 and human leukocyte antigen and negative for CD3, B lymphocyte antigen 36 and cytokeratin. Histopathological and immunohistochemical examinations confirmed a diagnosis of disseminated histiocytic sarcoma. This neoplasm has not been previously reported in a sugar glider.

Dystrophin, encoded by the DMD gene on the X chromosome, stabilizes the sarcolemma by linking the actin cytoskeleton with the dystrophin-glycoprotein complex (DGC). In-frame mutations in DMD cause a milder form of X-linked muscular dystrophy, called Becker muscular dystrophy (BMD), characterized by the reduced expression of truncated dystrophin. So far, no animal model with in-frame mutations in Dmd has been established. As a result, the effect of in-frame mutations on the dystrophin expression profile and disease progression of BMD remains unclear. In this study, we established a novel rat model carrying in-frame Dmd gene mutations (IF rats) and evaluated the pathology. We found that IF rats exhibited reduced expression of truncated dystrophin in a proteasome-independent manner. This abnormal dystrophin expression caused dystrophic changes in muscle tissues but did not lead to functional deficiency. We also found that the expression of additional dystrophin named dpX, which forms the DGC in the sarcolemma, was associated with the appearance of truncated dystrophin. In conclusion, the outcomes of this study contribute to the further understanding of BMD pathology and help elucidate the efficiency of dystrophin recovery treatments in Duchenne muscular dystrophy, a more severe form of X-linked muscular dystrophy., Editor's choice: The newly established rat model carrying in-frame mutations in the Dmd gene exhibits the dystrophic phenotype and abnormal dystrophin expression profile, similar to patients with Becker muscular dystrophy.

A 4-year and 10-month old female Pembroke Welsh Corgi presented with an enlarged right popliteal lymph node, and a histopathological diagnosis of nodal marginal zone lymphoma (nMZL) was made. After resection of the lymph node, follow-up observation was continued without chemotherapy. At 22 months after initial presentation, the dog developed enlargement of peripheral lymph nodes, and the histopathological diagnosis was late-stage nMZL. Multidrug chemotherapy induced clinical complete remission, but the tumor relapsed with enlargement of peripheral and abdominal lymph nodes 42 months after initial presentation. Second-round multidrug chemotherapy induced complete clinical remission again; however, the tumor relapsed with lymphadenopathy 47 months after initial presentation. The dog died 59 months after initial presentation, and postmortem examination revealed generalized lymphadenopathy; the histopathological diagnosis was diffuse large B-cell lymphoma (DLBCL). Polymerase chain reaction for antigen receptor gene rearrangements revealed that the nMZL and DLBCL samples were derived from the same B-lymphocyte clone.

Canine Lafora disease (LD) is an autosomal recessive genetic disorder causing nonfatal structural epilepsy, mainly affecting miniature wirehaired dachshunds. Repeat expansion in the EPM2B gene causes a functional impairment of the ubiquitin ligase malin which regulates glycogen metabolism. Abnormally structured glycogen accumulates and develop polyglucosan bodies predominantly in the central nervous system. The authors performed a comprehensive clinical, genetic, and pathological study of 4 LD cases affecting miniature wirehaired dachshund dogs with EPM2B repeat expansions, with systemic distribution of polyglucosan bodies and accumulation of laforin and other functionally associated proteins in the polyglucosan bodies. Myoclonic seizures first appeared at 7–9 years of age, and the dogs died at 14–16 years of age. Immunohistochemistry for calbindin revealed that the polyglucosan bodies were located in the cell bodies and dendritic processes of Purkinje cells. Polyglucosan bodies were also positive for laforin, hsp70, α/β-synuclein, ubiquitin, LC3, and p62. Laforin-positive polyglucosan bodies were located in neurofilament-positive neurons but not in GFAP-positive astrocytes. In nonneural tissues, periodic acid-Schiff (PAS)-positive polyglucosan bodies were observed in the heart, skeletal muscle, liver, apocrine sweat gland, and smooth muscle layer of the urinary bladder. In the skeletal muscle, polyglucosan bodies were observed only in type 1 fibers and not in type 2 fibers. The results indicate that although the repeat expansion of the EPM2B gene is specific to dogs, the immunohistochemical properties of polyglucosan body in canine LD are comparable to human LD. However, important phenotypic variations exist between the 2 species including the affected skeletal muscle fiber type.

A 12-year-old intact male Welsh Corgi was presented with enlargement of the right scrotum. Both testicles were surgically removed and histopathologically examined. On gross examination, white nodules were found in the epididymis and ductus deferens. Histopathologically, the nodules developed continuously from the tunica vaginalis testis of the right scrotum and consisted of spindle-shaped neoplastic cells that invaded the surrounding tissue. Immunohistochemically, the neoplastic cells were diffusely positive for vimentin, cytokeratin and Wilms tumor-1 (WT-1). Based on these findings, the tumor was diagnosed as sarcomatoid mesothelioma. The dog presented with respiratory distress 122 days after surgery and clinical examination found multiple metastatic lesions in the lung, abdominal lymph nodes and peritoneum. The dog died 144 days after surgery due to disease progression.

An 8-year-old male Japanese Shiba exhibited muscle wasting and a stiff gait. A low-amplitude myotonic discharge was recorded by needle electromyography (EMG). A histopathological examination on a tru-cut biopsy sample from the muscle revealed myofiber size variations. Internal nuclei and cytoplasmic vacuoles were observed in many fibers. A type 1 fiber predominance and many hybrid type fibers were observed immunohistochemically. On the basis of these EMG and histopathological findings, myotonic dystrophy (DM) was suspected as tentative diagnosis. The cytoplasm around the vacuoles was immunopositive for cytochrome c, tom 20, and SOD-1, suggesting that these vacuoles might occur within mitochondria. Collectively, these results indicate that a mitochondrial abnormality partly play the role on the pathogenesis of present case.

Primary epithelial tumors of the gallbladder are rarely reported in animals. In this study, 9 aged pigs (6–12 years old) were histopathologically examined for gallbladder proliferative lesions. At necropsy, a large gallstone occupied the lumen of the gallbladder of 3 pigs. Histopathological examination revealed chronic cholecystitis in all 9 pigs, mucosal hyperplasia in 2 pigs, adenoma in 1 pig, and adenocarcinoma in 2 pigs. Bacilli were detected in the gallbladder lumen of 6 pigs by Warthin-Starry stain. Mucosal hyperplasia, adenoma, and adenocarcinoma were characterized by papillary projections of the mucosa with occasional acinar structures. Tumor invasion of the surrounding tissue was observed in the cases of adenocarcinoma. On Alcian blue and periodic acid–Schiff double-stained sections, the acinar structure of gallbladder mucosa in chronic cholecystitis and mucosal hyperplasia was stained in a mosaic pattern, indicating pyloric gland metaplasia. The results of immunohistochemistry revealed a CD10-positive epithelial brush border and mucin (MUC) 2–positive goblet cells in chronic cholecystitis, adenoma, and adenocarcinomas, indicating intestinal metaplasia. Immunoreactivity of MUC5 AC and cytokeratin 19 was weaker in adenoma and adenocarcinomas compared with the normal and hyperplastic gallbladder mucosa. The number of p53-positive nuclei and the Ki-67 index were higher in adenocarcinomas compared with benign lesions. These results suggest that chronic cholecystitis associated with gallstones and/or bacterial infections may contribute to metaplastic changes and development of gallbladder tumors in aged pigs. Alteration of mucin, cytokeratin, and p53 profiles in gallbladder proliferative lesions in pigs were similar to that in humans, suggesting a common pathogenesis in tumor development.

Uteri from 50 four-toed hedgehogs ( Atelerix albiventris) with clinical signs of uterine disease were histopathologically examined. Sixteen animals (32%) were diagnosed with endometrial hyperplasia, 7 animals (14%) were diagnosed with endometrial polyp, and 27 animals (54%) were diagnosed with endometrial neoplasia. The mean ages of the animals with endometrial hyperplasia, polyp, and neoplasia were 28.7 months, 29.4 months, and 25.2 months, respectively. The neoplasms were classified into 7 endometrial mixed tumors, 12 endometrial stromal nodules, and 8 endometrial stromal sarcomas. However, the endometrial stromal nodules and endometrial stromal sarcomas often developed within or were contiguous with an endometrial polyp or mixed tumor. Interestingly, the stromal tumors and the stromal components of the endometrial polyp and mixed tumor displayed extraendometrial differentiation (eg, into adipocytes, granular cells, smooth muscle cells, and osteoid tissue). The endometrial stromal sarcomas exhibited severe cellular atypia and invaded subendometrial tissue. Immunohistochemical examinations demonstrated that the stromal cells of the hyperplastic lesions as well as the neoplastic lesions were positive for CD10, the progesterone receptor, and Wilms tumor 1. The four-toed hedgehog develops unique uterine neoplasms that are mainly composed of endometrial stromal cells and probably arise from endometrial polyps and/or mixed tumors.

Histochemical techniques used in examination of muscle biopsies typically require frozen sections. Given that most of the specimens submitted to a veterinary laboratory for diagnosis are formalin-fixed, the choice of staining methods is limited. We aimed to further advance the diagnostic capabilities of pathologists presented with formalin-fixed muscle samples and to describe the differences in immunohistopathologic findings between neurogenic and myogenic muscle disorders. Based on hematoxylin and eosin staining, we defined in dogs the histologic lesions in 4 neurogenic disorders (degenerative myelopathy and polyneuropathy) and 2 myogenic disorders (dystrophin-deficient muscular dystrophy). In cats, we defined the lesions in 2 neurogenic disorders (lymphoma of nerve roots and spinal cords) and 1 myogenic disorder (laminin α2–deficient muscular dystrophy). Immunohistochemistry for slow and fast myosins revealed angular and group atrophy of type 1 and type 2 fibers in dogs and cats, and fiber type grouping in dogs. These immunohistopathologic findings were specific to neurogenic muscle disorders. Immunohistochemistry for nestin and myogenin revealed nestin-positive fibers and myogenin-positive nuclei in dogs and cats. They were not specific, but these fibers in myogenic disorders can be interpreted as regenerating fibers. The immunohistochemical method described herein appears to be useful for discriminating neurogenic and myogenic disorders in formalin-fixed, paraffin-embedded muscle tissue of dogs and cats.

Sulfonylureas are often used alone or in combination with other drugs for treating type 2 diabetes. Ipragliflozin is a sodium/glucose cotransporter 2 inhibitor that enhances urinary glucose excretion and improves glycemic control. We examined the efficacy and safety of ipragliflozin as an add-on to a sulfonylurea in Japanese type 2 diabetes patients with inadequate glycemic control in a phase III study. Patients were randomized, double-blind, to 50 mg ipragliflozin or placebo for 24 weeks, followed by a 28-week open-label extension in which all patients received either 50 or 100 mg ipragliflozin. The primary endpoint was the change in HbA1c from baseline to week 24 (last observation carried forward). Overall, 166 patients were prescribed ipragliflozin and 77 were prescribed placebo. The adjusted mean change in HbA1c from baseline to week 24 between the ipragliflozin and placebo groups was −1.14 % (P < 0.001). The reductions in fasting plasma glucose and body weight were significantly greater in the ipragliflozin group (placebo-adjusted mean change: −38.0 mg/dl and −1.32 kg, respectively; both, P < 0.001). These changes were maintained until the end of the open-label extension. There were more treatment-emergent adverse events in the ipragliflozin group than in the placebo group (75.9 vs. 61.8 %; P = 0.032). The most common adverse events with a higher incidence in the ipragliflozin group than in the placebo group were mild pollakiuria and thirst. Ipragliflozin as an add-on to a sulfonylurea significantly improved glycemic control and reduced body weight, with good tolerability, in Japanese type 2 diabetes patients.

Ipragliflozin is a novel oral sodium–glucose cotransporter 2 (SGLT2) inhibitor under development for the treatment of type 2 diabetes. We examined its efficacy and safety as an add-on to pioglitazone in Japanese patients with inadequately controlled diabetes. Japanese type 2 diabetes patients were randomized to 24 weeks of treatment with 50 mg ipragliflozin or placebo in a double-blind manner. At week 24, patients with hemoglobin (Hb)A1c

The solution structure of the growth factor receptor-bound protein 2 (Grb2) SH2 domain complexed with a high-affinity inhibitor containing a non-phosphorus phosphate mimetic within a macrocyclic platform was determined by nuclear magnetic resonance (NMR) spectroscopy. Unambiguous assignments of the bound inhibitor and intermolecular NOEs between the Grb2 SH2 domain and the inhibitor was accomplished using perdeuterated Grb2 SH2 protein. The well-defined solution structure of the complex was obtained and compared to those by X-ray crystallography. Since the crystal structure of the Grb2 SH2 domain formed a domain-swapped dimer and several inhibitors were bound to a hinge region, there were appreciable differences between the solution and crystal structures. Based on the binding interactions between the inhibitor and the Grb2 SH2 domain in solution, we proposed a design of second-generation inhibitors that could be expected to have higher affinity.