Amputation of the Front Medial Digit of a Holstein Cow - Successful Parturition and Milk Production

Background: Diseases of the bovine digit and hoof remain major problems in dairy farming and the beef cow industry. Severe claudication accompanying swelling and pain at the lesion is often observed in deep digital sepsis, septic arthritis, severe sole ulcers, and chronic interdigital phlegmon. In addition, digital amputation is often performed in cases of white line disease or severe trauma, such as bone fractures. There are few reports of amputation of the medial digit of the bovine front hoof. In this case, amputation of the medial digit of the front leg of a pregnant cow was performed; its hospitalizationand return to productivity is reported.Case: The bovine was a 9-year-old Holstein cow in the 6th month of pregnancy, weighing 671 kg. Upon admission, the cow could not bear weight on its right front leg, and swelling and a purulent discharge were observed in the coronary area of the medial digit. X-ray examination results indicated a periosteal reaction centered on the 3rd phalanx of the medial digit of its right front leg hoof and slight periosteal reactions at the adaxial sides of the middle and proximal phalanges, strongly suggestive of septic arthritis caused by infection. According to the X-ray examination results, no abnormalities of the lateral digit of the front right hoof were observed; thus, it was estimated that the post-amputation hoof would be able to bear weight. Therefore, a decision was made to perform immediate amputation of the medial digit of the right front leg hoof to reduce pain for improved delivery of the offspring and improved milk production during the lactation period, rather than allowing the lesion progress until the dry period or the expected date of delivery. According to the X-ray examination results, amputation of the 1st phalanx alone was expected to suffice for removal of the cause of the pain; thus, a decision was made to perform amputation at the edge adjacent to the 2nd phalanx. The right front medial digit was prepared for aseptic surgery, and infiltration anesthesia was performed. An incision was made with a surgical knife at the interdigit of the right front limb. A wire saw was inserted into the site to cut the 2nd phalanx in the anti-axial direction down to the skin to resect the lesion. White viscous pus was discharged at the resection site; therefore, the wound was lavaged with tap water and packed with povidone iodine-impregnated gauze. The wound was also covered with a diaper and dressed with non-elastic and elastic bandages. The dressing was changed daily to aid wound healing. No problems were observed in the standing-up motion or other relevant movements immediately after the surgery. Granulation tissue formed rapidly, approximately 2 weeks after the surgery, and the lesion dried gradually. Approximately one month after the surgery, the subject exhibited little difficulty in both standing up and walking. The subject successfully delivered its 7th offspring at the farm on postoperative day 93.Discussion: The animal of this study was a 9-year-old, pregnant cow, and although amputation of the front medial digit is a relatively rare procedure, the cow was able to deliver and return to production, owing to sufficient postoperative treatment and care. The case also demonstrated the advantage of X-ray examination in bovine hoof diseases for accurate diagnosis, precise operation, and prognostic assessment. Keywords: bovine, lameness, surgery, septic arthritis, bovine hoof, medial digit, digital amputation

Resminostat in EGFR-mutated lung cancer

Drug-tolerant cells are mediators of acquired resistance. BIM-intron2 deletion polymorphism (BIM-del) is one of the mechanisms underlying the resistance to epidermal growth factor tyrosine kinase inhibitor (EGFR-TKI)-mediated apoptosis that induces drug tolerance. Here, we investigated whether resminostat, a histone deacetylase inhibitor, circumvents BIM-del-associated apoptosis resistance. The human EGFR-mutated non-small cell lung cancer (NSCLC) cell line PC-9 and its homozygous BIM-del-positive variant (PC-9 BIMi2-/-), established by editing with zinc finger nuclease, were used. In comparison with PC-9 cells, PC-9 BIMi2-/- cells were less sensitive to apoptosis mediated by EGFR-TKIs such as gefitinib and osimertinib. The combined use of resminostat and an EGFR-TKI preferentially induced the expression of the pro-apoptotic BIM transcript containing exon 4 rather than that containing exon 3, increased the level of pro-apoptotic BIM protein (BIMEL), and stimulated apoptosis in vitro. In a subcutaneous tumor model derived from PC-9 BIMi2-/- cells, gefitinib monotherapy decreased tumor size but retained residual lesions, indicative of the presence of tolerant cells in tumors. The combined use of resminostat and gefitinib increased BIMEL protein level and induced apoptosis, subsequently leading to the remarkable shrinkage of tumor. These findings suggest the potential of resminostat to circumvent tolerance to EGFR-TKIs associated with BIM deletion polymorphism

Podoplanin promotes progression of MPM

Malignant pleural mesothelioma (MPM) is characterized by dissemination and aggressive growth in the thoracic cavity. Podoplanin (PDPN) is an established diagnostic marker for MPM, but the function of PDPN in MPM is not fully understood. The purpose of this study was to determine the pathogenetic function of PDPN in MPM. Forty-seven of 52 tumors (90%) from Japanese patients with MPM and 3/6 (50%) MPM cell lines tested positive for PDPN. Knocking down PDPN in PDPN-high expressing MPM cells resulted in decreased cell motility. In contrast, overexpression of PDPN in PDPN-low expressing MPM cells enhanced cell motility. PDPN stimulated motility was mediated by activation of the RhoA/ROCK pathway. Moreover, knocking down PDPN with short hairpin (sh) RNA in PDPN-high expressing MPM cells resulted in decreased development of a thoracic tumor in mice with severe combined immune deficiency (SCID). In sharp contrast, transfection of PDPN in PDPN-low expressing MPM cells resulted in an increase in the number of Ki-67-positive proliferating tumor cells and it promoted progression of a thoracic tumor in SCID mice. Interestingly, PDPN promoted focus formation in vitro, and a low level of E-cadherin expression and YAP1 activation was observed in PDPN-high MPM tumors. These findings indicate that PDPN is a diagnostic marker as well as a pathogenetic regulator that promotes MPM progression by increasing cell motility and inducing focus formation. Therefore, PDPN might be a pathogenetic determinant of MPM dissemination and aggressive growth and may thus be an ideal therapeutic target

High efficacy of third generation EGFR inhibitor AZD9291 in a leptomeningeal carcinomatosis model with EGFR-mutant lung cancer cells

Leptomeningeal carcinomatosis (LMC) remarkably decreases the quality of life of EGFR-mutant lung cancer patients. In contrast to the lesions outside the central nervous system (CNS), molecular mechanisms of EGFR tyrosine kinase inhibitor (TKI) resistance in CNS lesions including LMC are largely unknown. In this study, we established an in vivo imaging model for LMC with EGFR mutant lung cancer cell lines harboring an exon 19 deletion in EGFR and evaluated the effect of first generation EGFR-TKIs, erlotinib, second generation afatinib, and third generation AZD9291. In PC-9/ffluc model, erlotinib treatment slowed the development of LMC. Importantly, treatment with afatinib or AZD9291 apparently delayed the development of LMC. Moreover, treatment with a higher dose of AZD9291, also associated with inhibited phosphorylation of EGFR downstream molecule S6, regressed LMC refractory to the aforementioned EGFR-TKI treatments. These observations suggest that the third generation EGFR-TKI AZD9291 may be an effective treatment for first or second generation EGFR-TKI resistant LMC caused by EGFR-mutant lung cancer

新生豚にみられた先天性過骨症

先天性過骨症は，前肢の硬化・腫脹を特徴とする新生豚の稀な疾患である。本疾患は常染色体の劣性遺伝によるものと考えられているが，原因や発生機序は明らかにされていない。病変部は四肢に限局し，橈骨・尺骨で顕著である。我々は分娩直後に死亡した雌の子豚に前肢の硬化・腫脹を認め，病理学的に先天性過骨症と診断した。組織学的に腫脹していた橈骨・尺骨では線維性骨梁が皮質骨表面から放射状に伸長し，周囲骨格筋は変性・萎縮し，線維化していた。後肢やその他臓器には著変はみられなかった。鑑別診断として，先天性筋症と肺性肥大性骨症が挙げられるが，前者は本例の病変が骨に主座し筋原線維の一次性構造異常がないことから，また後者は造骨形態や胸腔内に腫瘤病変がないことから否定された。Congenital hyperostosis is a rare disease of newborn pigs, characterized by thickening and sclerosis of the forelimbs. It has been suspected that it is inherited as an autosomal recessive trait, but the pathogenesis of the bone abnormalities has not been clarified. The lesions are characteristically localized in the limbs, and the radius and ulna are the most severely affected. We examined a female piglet who died right after delivery; the piglet showed thickening and sclerosis of the forelimbs, and the bone lesions were diagnosed as congenital hyperostosis. Histologically, fibrous trabecular bone proliferated radially and extended out from the surface of cortical bone of the swollen radius and ulna. The skeletal muscles around the bones showed degeneration and atrophy with fibrosis. No lesions were found in other bones and organs. For the differential diagnosis, congenital myopathy and pulmonary hypertrophic osteopathy were considered, but myopathy was excluded because of the lesion distribution and the absence of a primary structural abnormality of myofibrils, while pulmonary hypertrophic osteopathy was also excluded because of the different form of osteogenesis and the absence of a tumor in the thoracic cavity

Podoplanin promotes progression of malignant pleural mesothelioma by regulating motility and focus formation

Malignant pleural mesothelioma (MPM) is characterized by dissemination and aggressive growth in the thoracic cavity. Podoplanin (PDPN) is an established diagnostic marker for MPM, but the function of PDPN in MPM is not fully understood. The purpose of this study was to determine the pathogenetic function of PDPN in MPM. Forty-seven of 52 tumors (90%) from Japanese patients with MPM and 3/6 (50%) MPM cell lines tested positive for PDPN. Knocking down PDPN in PDPN-high expressing MPM cells resulted in decreased cell motility. In contrast, overexpression of PDPN in PDPN-low expressing MPM cells enhanced cell motility. PDPN stimulated motility was mediated by activation of the RhoA/ROCK pathway. Moreover, knocking down PDPN with short hairpin (sh) RNA in PDPN-high expressing MPM cells resulted in decreased development of a thoracic tumor in mice with severe combined immune deficiency (SCID). In sharp contrast, transfection of PDPN in PDPN-low expressing MPM cells resulted in an increase in the number of Ki-67-positive proliferating tumor cells and it promoted progression of a thoracic tumor in SCID mice. Interestingly, PDPN promoted focus formation in vitro, and a low level of E-cadherin expression and YAP1 activation was observed in PDPN-high MPM tumors. These findings indicate that PDPN is a diagnostic marker as well as a pathogenetic regulator that promotes MPM progression by increasing cell motility and inducing focus formation. Therefore, PDPN might be a pathogenetic determinant of MPM dissemination and aggressive growth and may thus be an ideal therapeutic target. © 2017 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association

Transient IGF-1R inhibition combined with osimertinib eradicates AXL-low expressing EGFR mutated lung cancer

Drug tolerance is the basis for acquired resistance to epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) including osimertinib, through mechanisms that still remain unclear. Here, we show that while AXL-low expressing EGFR mutated lung cancer (EGFRmut-LC) cells are more sensitive to osimertinib than AXL-high expressing EGFRmut-LC cells, a small population emerge osimertinib tolerance. The tolerance is mediated by the increased expression and phosphorylation of insulin-like growth factor-1 receptor (IGF-1R), caused by the induction of its transcription factor FOXA1. IGF-1R maintains association with EGFR and adaptor proteins, including Gab1 and IRS1, in the presence of osimertinib and restores the survival signal. In AXL-low-expressing EGFRmut-LC cell-derived xenograft and patient-derived xenograft models, transient IGF-1R inhibition combined with continuous osimertinib treatment could eradicate tumors and prevent regrowth even after the cessation of osimertinib. These results indicate that optimal inhibition of tolerant signals combined with osimertinib may dramatically improve the outcome of EGFRmut-LC

Allogeneic Hematopoietic Cell Transplantation for Blastic Plasmacytoid Dendritic Cell Neoplasm: A CIBMTR Analysis

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare hematological malignancy with a poor prognosis and considered incurable with conventional chemotherapy. Small observational studies reported allogeneic hematopoietic cell transplantation (allo-HCT) offers durable remissions in patients with BPDCN. We report an analysis of patients with BPDCN who received an allo-HCT, using data reported to the Center for International Blood and Marrow Transplant Research (CIBMTR). We identified 164 patients with BPDCN from 78 centers who underwent allo-HCT between 2007 and 2018. The 5-year overall survival (OS), disease-free survival (DFS), relapse, and nonrelapse mortality (NRM) rates were 51.2% (95% confidence interval [CI], 42.5-59.8), 44.4% (95% CI, 36.2-52.8), 32.2% (95% CI, 24.7-40.3), and 23.3% (95% CI, 16.9-30.4), respectively. Disease relapse was the most common cause of death. On multivariate analyses, age of ≥60 years was predictive for inferior OS (hazard ratio [HR], 2.16; 95% CI, 1.35-3.46; P = .001), and higher NRM (HR, 2.19; 95% CI, 1.13-4.22; P = .02). Remission status at time of allo-HCT (CR2/primary induction failure/relapse vs CR1) was predictive of inferior OS (HR, 1.87; 95% CI, 1.14-3.06; P = .01) and DFS (HR, 1.75; 95% CI, 1.11-2.76; P = .02). Use of myeloablative conditioning with total body irradiation (MAC-TBI) was predictive of improved DFS and reduced relapse risk. Allo-HCT is effective in providing durable remissions and long-term survival in BPDCN. Younger age and allo-HCT in CR1 predicted for improved survival, whereas MAC-TBI predicted for less relapse and improved DFS. Novel strategies incorporating allo-HCT are needed to further improve outcomes