A cDNA that encodes a novel Na ؉ -independent neutral amino acid transporter was isolated from FLC4 human hepatocarcinoma cells by expression cloning. When expressed in Xenopus oocytes, the encoded protein designated LAT3 (L-type amino acid transporter 3) transported neutral amino acids such as L-leucine, L-isoleucine, L-valine, and L-phenylalanine. The LAT3-mediated transport was Na ؉ -independent and inhibited by 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid, consistent with the properties of system L. Distinct from already known system L transporters LAT1 and LAT2, which form heterodimeric complex with 4F2 heavy chain, LAT3 was functional by itself in Xenopus oocytes. The deduced amino acid sequence of LAT3 was identical to the gene product of POV1 reported as a prostate cancer-upregulated gene whose function was not determined, whereas it did not exhibit significant similarity to already identified transporters. The Eadie-Hofstee plots of LAT3-mediated transport were curvilinear, whereas the low affinity component is predominant at physiological plasma amino acid concentration. In addition to amino acid substrates, LAT3 recognized amino acid alcohols. The transport of L-leucine was electroneutral and mediated by a facilitated diffusion. In contrast, L-leucinol, Lvalinol, and L-phenylalaninol, which have a net positive charge induced inward currents under voltage clamp, suggesting these compounds are transported by LAT3. LAT3-mediated transport was inhibited by the pretreatment with N-ethylmaleimide, consistent with the property of system L2 originally characterized in hepatocyte primary culture. Based on the substrate selectivity, affinity, and N-ethylmaleimide sensitivity, LAT3 is proposed to be a transporter subserving system L2. LAT3 should denote a new family of organic solute transporters.

System L is a major nutrient transport system responsible for the Na(+)-independent transport of large neutral amino acids including several essential amino acids. In malignant tumors, a system L transporter L-type amino acid transporter 1 (LAT1) is up-regulated to support tumor cell growth. LAT1 is also essential for the permeation of amino acids and amino acid-related drugs through the blood-brain barrier. To search for in vitro assay systems to examine the interaction of chemical compounds with LAT1, we have investigated the expression of system L transporters and the properties of [14C]L-leucine transport in T24 human bladder carcinoma cells. Northern blot, real-time quantitative PCR and immunofluorescence analyses have reveled that T24 cells express LAT1 in the plasma membrane together with its associating protein 4F2hc, whereas T24 cells do not express the other system L isoform LAT2. The uptake of [14C]L-leucine by T24 cells is Na(+)-independent and almost completely inhibited by system L selective inhibitor BCH. The profiles of the inhibition of [14C]L-leucine uptake by amino acids and amino acid-related compounds in T24 cells are comparable with those for the LAT1 expressed in Xenopus oocytes. The majority of [14C]L-leucine uptake is, therefore, mediated by LAT1 in T24 cells. Consistent with LAT1 in Xenopus oocytes, the efflux of preloaded [14C]L-leucine is induced by extracellularly applied substrates of LAT1 in T24 cells. This efflux measurement has been proven to be more sensitive than that in Xenopus oocytes, because triiodothyronine, thyroxine and melphalan were able to induce the efflux of preloaded [14C]L-leucine in T24 cells, which was not detected for Xenopus oocyte expression system. T24 cell is, therefore, proposed to be an excellent tool to examine the interaction of chemical compounds with LAT1.

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feline lower urinary tract disease (fLUtD) is a common problem in cats. the objectives of the study were to determine the prevalence, clinical signs, and causes of fLUtD and the risk factors for fLUtD. The medical records of 3486 cats visiting Chiang Mai University Small Animal Veterinary Teaching Hospital (VTH) between November 2016 and October 2017 were reviewed. An age-matched casecontrol study was performed to determine the risk factors for FLUTD by comparing 78 cats with FLUTD and 78 clinically normal age-matched cats. For each animal, potential risk data were obtained from medical records and cat owner interviews; these were analysed for associations with fLUtD. Multivariable logistic regression analysis was performed to estimate the odds ratios and to adjust for expected confounding factors. the prevalence of fLUtD in cats visiting the chiang Mai University Veterinary Teaching Hospital was 2.2%. The most common clinical signs identified were urethral obstruction (55.1%) and haematuria (23.1%). The most common diagnoses were feline idiopathic cystitis (FIC) (57.7%) and urolithiasis (struvite) (18%). The multivariable logistic regression analysis results indicated that fLUtD was most likely to be diagnosed in castrated male cats. fic and urolithiasis were the most common diagnoses in cats with fLUtD, and male sex and castration increased the risk of fLUtD.Feline lower urinary tract disease (FLUTD), a common disease in cats, is a syndrome that affects the reproductive tract, urinary bladder or urethra. The clinical signs of FLUTD are dysuria, haematuria, stranguria, pollakiuria and periuria. The prevalence of FLUTD among cats that had visited animal hospitals and clinics was 1.5% in the United States 1 and 2.2% in Bangkok, Thailand 2 . FLUTD can be obstructive or non-obstructive. Obstructions may occur as inflammatory debris from feline idiopathic cystitis (FIC), urethral plugs or uroliths. Non-obstructive causes include FIC, uroliths, urothelial carcinoma, anatomic defects (such as urethral stricture) and urinary tract infection (UTI) 3 .The risk factors for FLUTD differ across countries due to geography, season, diets 4 and cats' lifestyle. In New Zealand, low activity and indoor lifestyles were identified as risk factors for FLUTD 5 . In Norway, FLUTD was commonly found in male cats with an indoor lifestyle 6 . In Belgium, indoor confinement was the common risk factor for FLUTD 7 . In Bangkok, Thailand, a diet of commercial dry food and overweight were found to increase the risk for FLUTD 2 . In Austria, overweight cats had an increased risk for FLUTD, but the type of diet was not associated with FLUTD 8 .The Small Animal Veterinary Teaching Hospital in Chiang Mai, Thailand, receives a substantial number of feline patients each year. No one has yet evaluated the risk factors associated with FLUTD in this region. Therefore, the objectives of the present study were to determine the prevalence, clinical signs, and causes of FLUTD and risk factors for FLUTD in cats in Chiang Mai, Thailand.

Background: Ticks and fleas are considered amongst the most important arthropod vectors of medical and veterinary concern due to their ability to transmit pathogens to a range of animal species including dogs, cats and humans. By sharing a common environment with humans, companion animal-associated parasitic arthropods may potentially transmit zoonotic vector-borne pathogens (VBPs). This study aimed to molecularly detect pathogens from ticks and fleas from companion dogs and cats in East and Southeast Asia. Methods: A total of 392 ticks and 248 fleas were collected from 401 infested animals (i.e. 271 dogs and 130 cats) from China, Taiwan, Indonesia, Malaysia, Singapore, Thailand, the Philippines and Vietnam, and molecularly screened for the presence of pathogens. Ticks were tested for Rickettsia spp., Anaplasma spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. while fleas were screened for the presence of Rickettsia spp. and Bartonella spp. Result: Of the 392 ticks tested, 37 (9.4%) scored positive for at least one pathogen with Hepatozoon canis being the most prevalent (5.4%), followed by Ehrlichia canis (1.8%), Babesia vogeli (1%), Anaplasma platys (0.8%) and Rickettsia spp. (1%) [including Rickettsia sp. (0.5%), Rickettsia asembonensis (0.3%) and Rickettsia felis (0.3%)]. Out of 248 fleas tested, 106 (42.7%) were harboring at least one pathogen with R. felis being the most common (19.4%), followed by Bartonella spp. (16.5%), Rickettsia asembonensis (10.9%) and "Candidatus Rickettsia senegalensis" (0.4%). Furthermore, 35 Rhipicephalus sanguineus ticks were subjected to phylogenetic analysis, of which 34 ticks belonged to the tropical and only one belonged to the temperate lineage (Rh. sanguineus (sensu stricto)).

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The global emergence of canine parvovirus type 2c (CPV-2c) has been well documented. In the present study, 139 rectal swab samples collected from diarrheic dogs living in Vientiane, Laos, in 2016 were tested for the presence of the canine parvovirus (CPV) VP2 gene by PCR. The results showed that 82.73% (115/139) of dogs were CPV positive by PCR. The partial VP2 gene was sequenced in 94 of the positive samples; 91 samples belonged to CPV-2c (426Glu) subtype, while 3 samples belonged to the CPV-2a (426Asn) subtype. Notably, phylogenetic analysis of amino acid sequences revealed a close relationship between Laotian isolates and novel Chinese CPV-2c isolates. In Laotian CPV isolates, aligned protein sequences indicated a high rate of residue substitutions at positions 305, 324, 345, 370, 375, and 426 in the GH loop. The mutation at residue 370 (Q370R), a single mutation, was characterized as a unique mutant residue specific to the Laotian CPV-2c variant.

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