ABSTRACT. To investigate in vitro differentiation of canine adipose tissue-derived stromal cells (ATSCs) into neuronal cells, ATSCs from celiac adipose tissue in clinically healthy beagle dogs were treated with 100 µM dibutyryl cyclic adenosine monophosphate (dbcAMP) and 125 µM isobuthylmethylxanthine (IBMX). ATSCs were morphologically changed into differentiated ATSCs from spindle-shaped cells to neuron-like cells with numerous processes after the treatment. Expression of neuron-specific enolase (NSE) as an early neuron specific marker protein was detected in both ATSCs and differentiated ATSCs, however diachronic increase of NSE expression was observed in differentiated ATSCs after the treatment with dbcAMP/IBMX. In addition, neurofilament-68 (NF-68) as an early to mature neuron specific marker protein was weakly expressed in differentiated ATSCs. Neuron specific glutamate and glucose transporter (EAAC1 and GLUT-3, respectively) mRNAs were strongly expressed in differentiated ATSCs compared with those in ATSCs, although glia specific glutamate transporter mRNA (GLT-1) was also detected in differentiated ATSCs. ATSCs can differentiate into early to mature neuronal cells and are candidate cells for autologous nerve regeneration therapy, although additional research is needed to examine functional characteristics of differentiated ATSCs. KEY WORDS: adipose tissue-derived stromal cell, canine, dbcAMP/IBMX, in vitro differentiation, neuronal cell.
ABSTRACT. A three-month-old male Bull Terrier was referred to the Animal Medical Centre, Nihon University with chief complaints of subacute emesis and lethargy. Severe leukocytosis, high CRP, hypercalcemia and hypochloremia were detected. Moreover, severe calcification of gingival mucosa and abdominal skin, and abnormalities of the skeletal system were discerned. Abdominal X-ray and endoscopic examination revealed ulcer and hemorrhage on the mucosal membrane of the stomach. This might have been due to injections of high dose vitamin D at 3 and 2 weeks ago by another practioner, according to the detailed history of medication. After two months, a gastrointestinal and skin disorder disappeared, although calcification of the stomach membranes remained and abnormality of th e skeletal system had worsened. Therefore, vitamin D should be carefully administrated to a puppy. KEY WORDS: puppy, severe calcification, vitamin D toxicosis.J. Vet. Med. Sci. 66(9): 1133-1135, 2004 It is not clear why calcification of mucocutaneous and gastrointestinal tissues occurs, but it is well known that high dose administration of vitamin D induces hypercalcemia. Because we had an opportunity to examine a puppy, which suffered from calcification of some tissues caused by high dose administration of vitamin D, we report it in this paper.A three-month-old, 3.5 kg, male Bull Terrier was referred to the Nihon University Animal Medical Centre with subacute emesis, unequable abdominal pain, skin ulcer, polyuria, polydipsia and lethargy. The slightly emaciated patient puppy had usually licked many kinds of metals and plastic substances in its surroundings. Physical examination revealed a severe calcification of the gingival mucosa and abdominal skin, and systemic skeletal abnormalities (Fig. 1). A diagnosis of cholecaliciferol toxicosis was suggested by serum biochemical abnormalities; profound hypercalcemia (observed Ca, 16.4 mg/dl; reference range of 7.9 to 12.2 mg/dl in our hospital for less than 6 month old puppies), lower normal phosphorus concentration (5.0 mg/dl; reference range of 5.1 to 10.4 mg/dl), BUN (29.6 mg/dl) and creatinine (0.5 mg/dl). Increased neutrophils in number and high CRP were found on blood examination. Urine analysis indicated hypercalciuria (>16 mg/dl). Moreover, the serum 1-alpha-25 OH 2 Vitamin D value of the puppy was high (51.1 pg/ml). Abdominal radiography revealed that the barium administered 4 days before, remained in the stomach and a part of the descending colon. Gastric folds were observed as stripe-shaped calcification on radiography (Fig. 2a). Endoscopic examination showed ulcerous lesions and punctate hemorrhage on the mucosal membrane of the stomach, and food block covered with barium (Fig. 2b).The puppy was treated with antibiotics (orbifloxacin, 15 mg/head, sid), gastrointestinal mucous membrane protectant (sucralfate, 200 mg/head, bid), H 2 receptor-blocker (famotidine, 5 mg/head, bid) and predonisolone (1.5 mg/head, sid). Calcification of the gingival mucosa had disappeared, and his serum calcium ...
Structural determination of adsorbed atoms on layered structures such as clay minerals is a complex subject. Radioactive cesium (Cs) is an important element for environmental conservation, so it is vital to understand its adsorption structure on clay. The nuclear magnetic resonance (NMR) parameters of 133 Cs, which can be determined from solid-state NMR experiments, are sensitive to the local neighboring structures of adsorbed Cs. However, determining the Cs positions from NMR data alone is difficult. This paper describes an approach for identifying the expected atomic positions on clay minerals by combining machine learning (ML) with experimentally observed chemical shifts. A linear ridge regression model for ML is constructed from the smooth overlap of atomic position descriptor and gauge-including projector augmented wave (GIPAW) ab initio data. The constructed ML model predicts the GIPAW data to within a 3 ppm root-mean-squared error. At this stage, the 133 Cs chemical shifts can be instantaneously calculated from the Cs positions on any clay layers using ML. The inverse analysis, which derives the atomic positions from experimentally observed chemical shifts, is developed from the ML model. The input data for the inverse analysis are the layer structure and the experimentally observed chemical shifts. The Cs positions for the targeted chemical shifts are then output. Inverse analysis is applied to montmorillonite, and the resultant Cs positions are found to be consistent with previous results (Ohkubo, T.; et al. J. Phys. Chem. A 2018, 122, 9326−9337). The Cs positions on saponite clay are also clarified from experimentally observed chemical shifts and inverse analysis.
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