Long-term treatment of allogeneic adipose-derived stem cells in a dog with rheumatoid arthritis

Background@#Although there are growing demands for stem cell-based therapy for companion animals in various diseases, a few clinical trials have been reported. Moreover, most of them are the results from only one or a few times of stem cell injection. @*Objectives@#The aim of this study is to describe a long-term treatment with allogeneic adipose-derived stem cells (ASCs) in a dog with rheumatoid arthritis (RA), which is a rare canine disease. @*Methods@#The dog with RA received intravascular injection of allogeneic ASCs derived from two healthy donors once a month for 11 months. To assess therapeutic effects of ASCs, orthopedic examination and clinical evaluation was performed. Cytokines of tumor necrosis factor-α and interleukin-6 in the plasma were measured using ELISA analysis. @*Results@#Despite this repeated and long-term administration of allogeneic ASCs, there were no side effects such as immunorejection responses or cell toxicity. The orthopedic examination score for the dog decreased after ASCs treatment, and the clinical condition of the dog and owner’s satisfaction were very good @*Conclusions@#Although ASCs has been suggested as one of the options for RA treatment because of its anti-inflammatory and immunosuppressive functions, it has never been used to treat RA in dogs. The present report describes a case of canine RA treated with allogeneic ASCs for long-term in which the dog showed clinical improvement without adverse effects.

Mesenchymal Stem Cell and MicroRNA Therapy of Musculoskeletal Diseases

The therapeutic effects of mesenchymal stem cells (MSCs) in musculoskeletal diseases (MSDs) have been verified in many human and animal studies. Although some tissues contain MSCs, the number of cells harvested from those tissues and rate of proliferation in vitro are not enough for continuous transplantation. In order to produce and maintain stable MSCs, many attempts are made to induce differentiation from pluripotent stem cells (iPSCs) into MSCs. In particular, it is also known that the paracrine action of stem cell-secreted factors could promote the regeneration and differentiation of target cells in damaged tissue. MicroRNAs (miRNAs), one of the secreted factors, are small non-coding RNAs that regulate the translation of a gene. It is known that miRNAs help communication between stem cells and their surrounding niches through exosomes to regulate the proliferation and differentiation of stem cells. While studies have so far been underway targeting therapeutic miRNAs of MSDs, studies on specific miRNAs secreted from MSCs are still minimal. Hence, our ultimate goal is to obtain sufficient amounts of exosomes from iPSC-MSCs and develop them into therapeutic agents, furthermore to select specific miRNAs and provide safe cell-free clinical setting as a cell-free status with purpose of delivering them to target cells. This review article focuses on stem cell therapy on MSDs, specific microRNAs regulating MSDs and updates on novel approaches.

Mesenchymal Stem Cell and MicroRNA Therapy of Musculoskeletal Diseases

The therapeutic effects of mesenchymal stem cells (MSCs) in musculoskeletal diseases (MSDs) have been verified in many human and animal studies. Although some tissues contain MSCs, the number of cells harvested from those tissues and rate of proliferation in vitro are not enough for continuous transplantation. In order to produce and maintain stable MSCs, many attempts are made to induce differentiation from pluripotent stem cells (iPSCs) into MSCs. In particular, it is also known that the paracrine action of stem cell-secreted factors could promote the regeneration and differentiation of target cells in damaged tissue. MicroRNAs (miRNAs), one of the secreted factors, are small non-coding RNAs that regulate the translation of a gene. It is known that miRNAs help communication between stem cells and their surrounding niches through exosomes to regulate the proliferation and differentiation of stem cells. While studies have so far been underway targeting therapeutic miRNAs of MSDs, studies on specific miRNAs secreted from MSCs are still minimal. Hence, our ultimate goal is to obtain sufficient amounts of exosomes from iPSC-MSCs and develop them into therapeutic agents, furthermore to select specific miRNAs and provide safe cell-free clinical setting as a cell-free status with purpose of delivering them to target cells. This review article focuses on stem cell therapy on MSDs, specific microRNAs regulating MSDs and updates on novel approaches.

Preputial gland adenoma in a wild nutria (Myocastor coypus): a case report

Tumor incidence in wild mammals is reportedly very low. Wild nutria, a large rodent, is known to carry many infectious diseases, but rarely exhibits neoplastic diseases. We necropsied a male wild nutria and found a large nodular mass in the left inguinal region, adjacent to the penis. Histopathologically, the mass was diagnosed as preputial gland adenoma. Spontaneous preputial gland adenomas are extremely rare in all animals. Moreover, reports of tumors in nutrias have been limited to adenocarcinomas of the lungs and uterus, as well as subcutaneous fibromas. Here, we describe preputial gland adenoma in a wild nutria.

Preputial gland adenoma in a wild nutria (Myocastor coypus): a case report

Tumor incidence in wild mammals is reportedly very low. Wild nutria, a large rodent, is known to carry many infectious diseases, but rarely exhibits neoplastic diseases. We necropsied a male wild nutria and found a large nodular mass in the left inguinal region, adjacent to the penis. Histopathologically, the mass was diagnosed as preputial gland adenoma. Spontaneous preputial gland adenomas are extremely rare in all animals. Moreover, reports of tumors in nutrias have been limited to adenocarcinomas of the lungs and uterus, as well as subcutaneous fibromas. Here, we describe preputial gland adenoma in a wild nutria.

Preputial gland adenoma in a wild nutria (Myocastor coypus): a case report

Pathomorphological features of calcinosis circumscripta in a dog / 한국실험동물학회지

A 19-month-old male German shepherd dog was presented with a hindlimb footpad mass. The mass was excised and histopathology was performed. Grossly, the cut section of the mass had multiple well-circumscribed nodules with a chalky appearance. Histopathologically, basophilic-stained calcium deposits of various sizes were distributed from dermis to subcutis, surrounded by epithelioid cells, multinucleated giant cells, and reactive fibroblasts. Myxoid metaplasia, calcium deposits in hair follicles, and psammoma-like bodies were also found. These histopathologic observations will greatly help to understand the pathogenesis of calcinosis circumscripta.

Loquat (Eriobotrya japonica) leaf extract inhibits the growth of MDA-MB-231 tumors in nude mouse xenografts and invasion of MDA-MB-231 cells

BACKGROUND/OBJECTIVES: The present study was conducted to examine the inhibitory effect of loquat leaves on MDA-MB-231 cell proliferation and invasion. MATERIALS/METHODS: Female athymic nude mice were given a subcutaneous (s.c.) inoculation of MDA-MB-231 cells and randomly grouped to receive a s.c. injection of either 500 mg/kg ethanol, water extract or vehicle five times a week. Tumor growth, mitotic rate and necrosis were examined. MDA-MB-231 cells were cultured with DMSO or with various concentrations of loquat water or ethanol extract. Proliferation, adhesion, migration, invasion and matrix metalloproteinase (MMP) activity were examined. RESULTS: Tumor growth of xenograft nude mouse was significantly reduced by loquat extracts. The results of mitotic examination revealed that loquat extracts reduced tumor cell division. Both ethanol and water extracts significantly inhibited MDA-MB-231 cell proliferation. The protein expression of ErbB3 was significantly down-regulated by loquat leaf extracts. Loquat leaf extracts increased apoptosis of MDA-MB-231 cells following 24 hour incubation and the ethanol extract was more potent in inducing apoptosis than the water extract. Furthermore, loquat extracts inhibited adhesion, migration and invasion of MDA-MB-231 cells. MMP activity was significantly inhibited by loquat extracts. CONCLUSION: Our results show that extracts of loquat inhibit the growth of tumor in MDA-MB-231 xenograft nude mice and the invasion of human breast cancer cells, indicating the inhibition of tumor cell proliferation and invasion.

St. John's Wort (Hypericum perforatum) stimulates human osteoblastic MG-63 cell proliferation and attenuates trabecular bone loss induced by ovariectomy

BACKGROUND/OBJECFTIVES: The effect of St. John's Wort extract (SJW) on MG-63 cell proliferation and trabecular bone loss induced by ovariectomy was examined. MATERIALS/METHODS: Proliferation, expression of estrogen receptor (ER) alpha and ER beta, and gene expressions of osteoprotegerin (OPG), osteocalcin (OC) and alkaline phosphatase (ALP) were examined in MG-63 cells treated with or without SJW. Ovariectomized rats were treated with SJW at the dose of 100 or 200 mg/kg/day, beta-estradiol-3-benzoate (E2), or vehicle only (OVX-C), and sham operated rats were treated with vehicle only (Sham-C). Serum ALP and C-telopeptide (CTX), and femoral trabecular bone loss were examined. RESULTS: SJW increased MG-63 cell proliferation and expression of ER alpha and ER beta, and positive effect was shown on gene expressions of ALP, OC and OPG. SJW also showed estrogen like effect on bone associated with slowing down in trabecular bone loss. Histopathology by H&E showed rats treated with SJW displayed denser structure in metaphyseal region of distal femur compared with rats in OVX-C. SJW was shown to reduce serum CTX in OVX rats. CONCLUSION: The present study provides new insight in preventing estrogen deficiency induced bone loss of SJW and possibility for its application in bone health supplement.

Effect of St. John's Wort (Hypericum perforatum) on obesity, lipid metabolism and uterine epithelial proliferation in ovariectomized rats

BACKGROUND/OBJECTIVES: This study was conducted to assess the potential of St. John's Wort (Hypericum perforatum) to prevent obesity and abnormalities in lipid metabolism induced by ovariectomy in a rat model without stimulatory activity on uterus. MATERIALS/METHODS: Ovariectomized (OVX) rats were treated for 6 weeks with 70% ethanol extracts of Hypericum perforatum [HPEs: whole plant (WHPE) and flower and leaves (FLHPE)], beta-estradiol-3-benzoate at a dose of 50 microg/kg/day (E2) or vehicle (distilled water). RESULTS: As expected, OVX increased body weight gain and adiposity and showed higher food efficacy ratio. OVX also increased the serum cholesterol as well as insulin resistance, while reducing uterus weight and uterine epithelial proliferation rate. HPEs (WHPE and FLHPE) showed estrogen-like effect on body weight gain, adipose tissue weight and food efficacy ratio in OVX rats. HPEs prevented hypercholesterolemia induced by OVX more effectively than E2. E2 increased uterus weight and epithelial proliferation rate in OVX rats, while HPEs maintained them at the level of the sham-operated animals. CONCLUSIONS: Our finding demonstrates that HPEs can be considered as an effective agent to prevent OVX-induced obesity without stimulatory activity on uterus.

Primary leiomyoma of the ovaries in a dog / 동물의과학연구지

We would like to report a case of leiomyoma of the ovaries in a dog. Leiomyoma is commonly seen in the vagina in dogs. However, it is a very rare neoplasm in the ovaries. As there have only been a few reported cases, this report provides valuable information on veterinary medicine and pathology. Masses found in the ovaries need to be differentiated from other ovarian tumors. Therefore, we describe the gross, histopathological, and immunohistochemical features of a case of ovarian leiomyoma in a 10-year-old female Yorkshire Terrier dog. The mass on the right of the uterus was found accidentally by pelvic ultrasonography. Laparatomy revealed a large multi-nodulated ovarian mass. Grossly, cut surfaces of the mass showed multiple firm whitish nodules in the cortex and bloody loose connective tissue in the medulla. Histopathologically, the cortex of the mass was composed of spindle cells forming interlacing fascicles. The cells had elongated, blunt-ended nuclei and eosinophilic cytoplasm as detected by hematoxylin and eosin staining. Immunohistochemical stained sections were immunoreactive for alpha-smooth muscle actin and desmin but negative for vimentin and S-100. Therefore, differential diagnosis confirmed leiomyoma based on morphology and positive staining for alpha-smooth muscle actin and desmin.

A case of metastatic cholangiocarcinoma in a ferret / 동물의과학연구지

Cholangiocarcinoma is a relatively rare neoplasm in animals and humans. A four-year-old, neutered male ferret presented with depression, anorexia, cachexia, diarrhea, and icterus. Necropsy findings included ascites, multiple white nodules on the surface of the liver, stomach, and duodenum, gross enlargement of the bile duct and right atrium, hemorrhage of the gastric and duodenal mucosa, and icterus of the mesenteric fat. Infiltrative well-differentiated neoplastic biliary epithelial cells forming ducts and acini with a prominent collagenous stroma were observed on microscopic examination of neoplastic lesions within the liver, mesentery, and the serosa of the stomach and duodenum. This is a report on a rare case of obstructive jaundice due to cholangiocarcinoma in a ferret.

Chronic pyometra due to bilateral ovarian tumors in a South American sea lion / 동물의과학연구지

A 14-year-old female South American sea lion (Otaria byronia) with persistent vaginal secretion and chronic hemorrhagic diarrhea was encountered. During postmortem examination, the uterus was found to resemble a balloon with mucosal congestion and was filled with grayish milky material. The ovaries also had abnormal features, including necrotic surface lesions and multiple whitish foci in the cut section. Hemorrhages and ulcerated changes due to toxemia were observed in other organs, including the liver, spleen, lung, intestines, and lymph nodes. Microscopically, the left ovary contained interlacing fascicles of fibroblast-like cells with blunt-end nuclei showing cytoplasmic positive immunoreactivity against alpha-smooth muscle actin and desmin. The right ovary contained cells with round to cigar-shaped nuclei showing cytoplasmic positive immunoreactivity against vimentin. In conclusion, based on classification of bilateral ovarian tumors as a leiomyoma in the left region and a fibroma in the right region, this sea lion was diagnosed with chronic closed pyometra.

Case of Sporadic T-cell Lymphosarcoma in Cattle / 동물의과학연구지

We would like to report a case of bovine lymphosarcoma. Parous cattle from a livestock farmhouse were examined for mutinodular masses in the abdominal cavity after slaughter. For clinical signs, animals presented mild leukemic signs but did not have viral or bacterial infection. Grossly, whitish to yellowish smooth masses similar to fat tissue were covered with a thin membrane. A multilobulated mass formed around the arteri, and there was a large quantity of reddish fluid on the cut surface. Histopathologically, a monomorphic population of lymphocytes was observed along with small amounts of cytoplasm, round nuclei with coarsely granular chromatin, and numerous mitotic figures in the samples. In the tumor lesion, uniformly round cells had invaded with abundant neovascularization. Especially, the immunohistochemical phenotype of tumor cells was positive for anti-CD3 and negative for anti-CD8 and anti-CD20. Therefore, morphological analysis diagnosed the mass as a multinodular bovine lymphosarcoma of T-cell origin without any sign of infection by a viral agent.

Angiotropic metastatic malignant melanoma in a canine mammary gland / 한국실험동물학회지

An eleven-year-old spayed female Yorkshire Terrier presented with a sublumbar mass and upon ultrasonographic examination, was revealed to have a mammary gland tumor. Black to reddish colored masses, located in the visceral peritoneum of the sublumbar region was observed on laparotomy with masectomy of the right side. In the laparotomy, we observed reddish masses multifocally located in the serosal membrane of the large intestine. Histopathologic examination of the intestinal and abdominal mass showed highly invasiveness into the muscle and metastasis of melanocytic tumor cells through the blood vessels. The mammary glands showed abnormal hyperplasia of melanocytes, destruction of the normal glands by tumor cells and infiltration of some lymphocytes in the pool of melanocytic cells. We have identified a malignant melanoma containing an angiotumoral complex in which tumor cells occupied a pericytic location along the microvessels with intravasation determined by immunohistochemistry for S100 protein and protein kinase C-alpha. Histologic findings in this dog lead to a diagnosis of an angiotropic metastatic malignant melanoma.

Canine hemangiosarcoma in a female Jindo dog / 한국실험동물학회지

A fourteen-year female Jindo dog showing signs of hemoperitoneum was diagnosed with hemangiosarcoma through abdominal radiography and histopathological examination. In the abdominal radiographs, a circular mass was observed on the posterior of the spleen. When the splenic mass was removed via laparotomy, it was spherical, poorly circumscribed, and showed signs of necrosis, with white spots present on the outer side of the mass. Microscopically, the mass showed that hemorrhage was widespread and extensive infiltration of neoplastic cells was found throughout. There was hypercellularity, including occasional pleomorphic cells and mitotic figures. Inflammation at the edge of the mass, along with necrosis, was also found. In this report, we describe the gross and histopathological findings of a case of canine hemangiosarcoma.

Pilomatrixoma in a Maltese Dog / 한국실험동물학회지

A 12-year-old female Maltese dog brought to a local veterinary clinic with two nodules in the subcutis of left shoulder. The nodules were grown gradually from 2 years ago. The nodules were removed surgically under anesthesia and examined histopathologically. They were 15x26 mm in diameter (oval form) and 12x15 mm in diameter (round form), respectively. Histopathologically, the lesions consisted of multilayered basaloid cells in the peripheral of the mass and ghost cells in the central region. Typical findings of these two nodules were gradually keratinized basaloid cell toward central area result in forming anuclear ghost cells. These microscopic features of cutaneous nodules were diagnosed as pilomatrixoma and the identified findings were similar to fully developed stage of human pilomatrixoma. This report may assist in the categorizing of canine pilomatrixoma using histopathological features.

Fractionation of DNases Specific to Haemonchus contortus Intestine by Phenyl Sepharose Column / 한국실험동물학회지

Multiple DNases were identified from Haemonchus contortus intestine based on previous studies. The DNases detected at 34, 36 and 38.5 kDa had diverse characteristics. Some of them had characteristics similar to those of mammalians and others had unusual characteristics. This study was carried out to fractionate worm intestinal DNases from other proteins using phenyl Sepharose chromatographic methods. All DNases detected from Haemonchus contortus intestine were fractionated in the flowthrough of phenyl Sepharose, indicating the worm DNases are hydrophilic. The DNases were enriched five-fold in the flowthrough fraction while additional steps are required for isolation of the worm DNases. Thus, fractionation with phenyl Sepharose could be used as a good initial step to enrich and separate DNases from other proteins.

Fractionation of DNases Specific to Haemonchus contortus Intestine by Phenyl Sepharose Column / 한국실험동물학회지

Multiple DNases were identified from Haemonchus contortus intestine based on previous studies. The DNases detected at 34, 36 and 38.5 kDa had diverse characteristics. Some of them had characteristics similar to those of mammalians and others had unusual characteristics. This study was carried out to fractionate worm intestinal DNases from other proteins using phenyl Sepharose chromatographic methods. All DNases detected from Haemonchus contortus intestine were fractionated in the flowthrough of phenyl Sepharose, indicating the worm DNases are hydrophilic. The DNases were enriched five-fold in the flowthrough fraction while additional steps are required for isolation of the worm DNases. Thus, fractionation with phenyl Sepharose could be used as a good initial step to enrich and separate DNases from other proteins.

Two different types of malignant fibrous histiocytomas from pet dogs

We describe 2 cases of malignant fibrous histiocytomas (MFHs) that spontaneously developed in young pet dogs. To classify these tumors, we applied a panel of antibodies (vimentin, desmin, alpha-SMA, and ED1) and Azan staining for collagen. The MFHs were most consistent with osteoclast-like giant and inflammatory cell types. The first case had positive staining for ED1 and vimentin, and given the osteoclast-like giant cells, calcification sites accompanying peripheral giant cell infiltrates. The latter case, the inflammatory cell type, exhibited a storiform-pleomorphic variant of neoplastic cells, including an ossifying matrix. MFHs are among the most highly aggressive tumors occurring in soft tissue sarcomas in elderly dogs; however, MFHs have been poorly studied from a diagnostic point of view. Herein, we describe the histologic and immunohistologic features of MFHs in detail, thus classifying the subtypes of these tumors.