Mutation of the Thap4 gene causes dwarfism and testicular anomalies in rats and mice.

The petit (pet) locus is associated with dwarfism, testicular anomalies, severe thymic hypoplasia, and high postnatal lethality, which are inherited in autosomal recessive mode of inheritance in rats with a Wistar strain genetic background. Linkage analysis localized the pet locus between 98.7 Mb and 101.2 Mb on rat chromosome 9. Nucleotide sequence analysis identified 2 bp deletion in exon 2 of the Thap4 gene as the causative mutation for pet. This deletion causes a frameshift and premature termination codon, resulting in a truncated THAP4 protein lacking approximately two-thirds of the C-terminal side. Thap4 is expressed in various organs, including the testis and thymus in rats. To elucidate the biological function of THAP4 in other species, we generated Thap4 knockout mice lacking exon 2 of the Thap4 gene through genome editing. Thap4 knockout mice also exhibited dwarfism and small testis but did not show high postnatal lethality. Thymus weights of adult Thap4 knockout male mice were significantly higher compared to wild-type male mice. Although Thap4 knockout male mice were fertile, their testis contained seminiferous tubules with spermatogenesis and degenerative seminiferous tubules lacking germ cells. Additionally, we observed vacuoles in seminiferous tubules, and clusters of cells in the lumen in seminiferous tubules in Thap4 knockout male mice. These results demonstrate that spontaneous mutation of Thap4 gene in rats and knockout of Thap4 gene in mice both cause dwarfism and testicular anomalies. Thap4 gene in rats and mice is essential for normal testicular development, maintaining spermatogenesis throughout the entire region of seminiferous tubules.

Molecular characterization of GPR84 in domestic cats.

G protein-coupled receptor 84 (GPR84) was cloned as an orphan receptor, and medium-chain fatty acids were then revealed as endogenous ligands. GPR84 is expressed in immune cells and is believed to protect liver function from lipotoxicity caused by overeating and high-fat diet intake. This study aimed to present the molecular characterization of GPR84 in domestic cats. The deduced amino acid sequence of the feline GPR84 shows high sequence homology (83-89 %) with the orthologues from other mammalians by cDNA cloning of feline GPR84. Remarkably high mRNA expression was observed in the bone marrow by Q-PCR analysis. The inhibition of intracellular cAMP concentration was observed in cells transfected with feline GPR84 and treated with medium-chain fatty acids. Immunostaining of GPR84 and free fatty acid receptor 2 (FFAR2)/GPR43 in the bone marrow, where high mRNA expression was observed, showed reactions in macrophages and myeloid cells. To clarify whether the receptor formed homo/hetero-merization, GPR84 and FFARs were analyzed using Nano-Luc binary technology and NanoLuc bioluminescence resonance energy transfer technologies, which revealed that GPR84 formed more heteromers with FFAR2 than homomers with each other. In addition, when GPR84 and FFAR2/GPR43 were cotransfected in the cell, their localization on the cell membrane was reduced compared with that when single receptors were transfected. These results indicated that GPR84 is a functional receptor protein that is expressed in cat tissues and may have a protein-protein interaction with FFAR2/GPR43 on the cell membrane.

The Physiological Function of Deglutition and Swallowing in Patients With Maxillary Protrusion After Mandibular Advancement.

The objective of this study was to determine the tongue-palatal contact changes in patients with skeletal maxillary protrusion after sagittal split ramus osteotomy (SSRO) during swallowing. In this study, 15 patients with maxillary protrusion and 10 normal subjects participated. Before and 3 months after surgery, tongue-palatal contact patterns during swallowing of patients with maxillary protrusion as well as controls were evaluated by electropalatography. The electrode contact number in the alveolar, palatal, and velar parts was examined. The swallowing duration of each phase was also evaluated. In the lateral area of the velar part, incomplete electrode contact was shown at 0.3 seconds in patients with maxillary protrusion. The electrode contact number in the velar part at 0.3 seconds before tongue-palatal complete contact was significantly less in the preoperative patients compared with the controls ( P < 0.05). A small increase in the electrode contact number of the velar part was shown in the postoperative patients at 0.3 and 0.2 seconds before tongue-palatal complete contact ( P < 0.05). The pharyngeal phase duration was significantly larger in the patients with maxillary protrusion before SSRO compared with the controls ( P < 0.05). After SSRO, the pharyngeal phase duration was significantly shortened. It was shown that the tongue-palatal contact pattern during swallowing in patients with maxillary protrusion improved after orthognathic surgery, and the pharyngeal phase duration was also shortened. It is suggested that the changes in the mesiodistal mandibular position by orthognathic surgery can improve tongue posture and movement during swallowing.

EPG research and therapy: further developments.

Electropalatography (EPG) has been used in the past 50 years for studying the patterns of contact between the tongue and the palate during speech production in typical speakers and those with speech disorders due to different causes. At the 7th EPG Symposium in Japan that was held online on 24 January 2021 (see: https://epg-research.sakura.ne.jp/), a panel of invited experts discussed their views regarding further developments and application of the technique. This paper provides a summary of this discussion. EPG offers information on articulation which cannot be replaced by other instrumental measures of speech. Identified areas for further hardware development are thinner EPG plates, better dental and palatal coverage, wireless connectivity, and sensors that provide additional articulatory information (e.g. tongue pressure, tongue-palate distance). EPG can serve as a resource for teaching speech disorders and phonetics. Furthermore, EPG therapy can be combined with telepractice in the speech therapy of clients with speech disorders.

Electropalatography (EPG) activities in Japan and the impact of the COVID-19 pandemic on EPG research and therapy: A report of presentations at the 7th EPG Symposium.

BACKGROUND: At the 7th Electropalatography Symposium in Japan, held online on the 24 January 2021, a few speakers were invited to talk about how the COVID-19 pandemic had impacted their research and/or speech therapy that involved the use of electropalatography (EPG) as well as the procedures adopted in order to continue their work in a safe manner. The information on protective measures when using instrumental techniques in speech research and therapy may be useful for colleagues in research and the clinic. AIMS: The primary aims are: (1) to find out whether there are any published recommendations regarding protective measures for using EPG in research and clinic settings; (2) to discuss the impact of the pandemic and the corresponding restrictions and general protective measures directed (or advised) by local government and professional bodies at each stage of EPG work; and (3) to share experiences in using modified procedures for face-to-face EPG therapy sessions and combined EPG teletherapy. In addition, a brief overview of EPG and a summary of EPG research and clinical activities in Japan presented by one of the symposium organizers at the symposium are included. METHODS & PROCEDURES: A review of the literature regarding protective measures recommended for using EPG for speech assessment and treatment or research, supplemented by a discussion of our own experiences. MAIN CONTRIBUTION: The literature review showed that there are no guidelines regarding protective measures for using EPG, but there is some advice regarding speech recording using microphones. Most published articles related to speech and language therapy (SLT) service during COVID-19 are about telepractice or general clinical guidelines for face-to-face speech therapy sessions. The protective measures for using EPG developed based on the general guidelines recommended by local government and professional bodies (e.g., using visors, transparent acrylic board) were described. Using EPG in telepractice was discussed as well. CONCLUSIONS: It has been challenging to continue EPG research and therapy during the pandemic. In order to deal with this crisis, available knowledge regarding infection control and recommendations from local government and professional bodies were applied to design methods and procedures that allowed EPG research and therapy to continue. WHAT THIS PAPER ADDS: What is already known on the subject There are general protective measures recommended by local government and professional bodies regarding speech therapy sessions (e.g., using personal protective equipment (PPE), social distancing), but little is known about the measures for using instrumental techniques in speech research and therapy, particularly EPG. The equipment of each instrumental technique is different, so measures that are appropriate for one may not be suitable for others. Hence, specific recommendations are needed for EPG. What this paper adds to existing knowledge This paper provides pointers to information about recommendations regarding protective measures for speech research and therapy, supplemented with suggestions specific to EPG provided by experienced users based on actual experience. What are the potential or actual clinical implications of this work? In evaluating the impact of the COVID-19 pandemic on EPG research and therapy, an analytical approach was taken to break down the steps involved in carrying out those activities, and the challenges we faced and the possible alternatives for completing the tasks were discussed. A similar approach can be applied to evaluate other aspects of speech therapy service.

An Investigation of Tongue-Palatal Contact Changes in Patients With Skeletal Mandibular Lateral Deviation After Sagittal Split Ramus Osteotomy.

ABSTRACT: This study aimed to investigate the changes in tongue-palatal contact patterns in patients with mandibular lateral deviation by electropalatography (EPG) before and after sagittal split ramus osteotomy (SSRO). Ten mandibular asymmetry patients who underwent SSRO participated in the study. Tongue-palatal contact patterns for the production of /t/ and /s/ sounds were observed using EPG before surgery and 3 months after surgery, and the changes in EPG pattern were examined. The number of electrode contacts in the 2 vertical columns of the EPG plate was calculated both in the mandibular deviation side and the nondeviation side. The EPG patterns for /t/ and /s/ showed asymmetry before surgery but became normal after surgery. Before surgery, the number of electrode contacts in the 2 vertical columns in the mandibular deviation side was significantly lower than that in the nondeviation side and the normal participants during /t/ and /s/ articulation. However, the number of electrode contacts in the deviation side significantly increased after surgery. This study demonstrated that the tongue-palatal contact patterns for /t/ and /s/ articulation shifted to the direction of mandibular deviation and improved after SSRO.

Association between chemical components of PM2.5 and children's primary care night-time visits due to asthma attacks: A case-crossover study.

BACKGROUND: Few papers have examined the association between the chemical components of PM2.5 and health effects. The existence of an association is now under discussion. METHODS: This case-crossover study aimed to examine the association between the chemical components of PM2.5 and night-time primary care visits (PCVs) due to asthma attacks. The subjects were 1251 children aged 0-14 years who received medical care for asthma at a municipal emergency clinic. We measured daily average concentrations of hydrogen ion, sulfate ion, nitrate ion and water-soluble organic compounds (WSOCs), which are components of PM2.5. We estimated the odds ratios (ORs) of PCVs per unit increment (inter quartile ranges) in each chemical component of PM2.5 for the subgroups of warmer months and colder months separately. RESULTS: No association was seen between PCVs and PM2.5 mass concentrations the day before the PCVs in either warmer or colder months. In the warmer months, an association was seen with the concentrations of WSOCs and hydrogen ion the day before the PCVs (OR = 1.33; 95% CI: 1.00-1.76, OR = 1.18; 95% CI: 1.02-1.36, respectively). Furthermore, a negative association was seen between sulfate ion and PCVs (OR = 0.85; 95%CI: 0.74-0.98). No associations were observed in the colder months. CONCLUSIONS: We observed a positive association between PCVs and certain concentrations of WSOCs and hydrogen ions in warmer months. In contrast, sulfate ion showed a negative association.

Molecular characterization of feline melanocortin 4 receptor and melanocortin 2 receptor accessory protein 2.

Melanocortin 4 receptor (MC4R), which is a member of the G protein-coupled receptor (GPCR) family, mediates regulation of energy homeostasis upon the binding of α-melanocyte-stimulating hormone (α-MSH) in the central nervous system (CNS). Melanocortin 2 receptor accessory protein 2 (MRAP2) modulates the function of MC4R. We performed cDNA cloning of cat MC4R and MRAP2 and characterized their amino acid sequences, mRNA expression patterns in cat tissues, protein-protein interactions, and functions. We found high sequence homology (>88%) with other mammalian MC4R and MRAP2 encoding 332 and 206 amino acid residues, respectively. Reverse transcription-polymerase chain reaction analysis revealed that cat MC4R and MRAP2 mRNA were expressed highly in the CNS. In CHO-K1 cells transfected with cat MC4R, stimulation with α-MSH increased intracellular cyclic adenosine monophosphate (cAMP) concentration in a dose-dependent manner. Furthermore, the presence of MRAP2 enhanced the cat MC4R-mediated cAMP production. These results suggested that cat MC4R acts as a neuronal mediator in the CNS and that its function is modulated by MRAP2. In addition, our NanoBiT study showed the dynamics of their interactions in living cells; stimulation with α-MSH slightly affected the interaction between MC4R and MRAP2, and did not affect MC4R homodimerization, suggesting that they interact in the basal state and that structural change of MC4R by activation may affect the interaction between MC4R and MRAP2.

Changes in fatty acid composition in tissue and serum of obese cats fed a high fat diet.

BACKGROUND: Obesity and overweight have been frequently observed in dogs and cats in recent years as in humans. The compositions of fatty acids (FAs) in the accumulated lipids in tissues of obese animals may have important roles in the process and mechanisms related to the onset of metabolic disorders. The purpose of this study was to evaluate the effects of a high fat (HF) diet, which contained a higher proportion of saturated FAs, on FA metabolism and distribution in obese cats. Cats (N = 12) were divided into control diet group (crude fat; 16.0 %) (n = 4) or a high fat (HF) diet group (crude fat; 23.9 %) (n = 8). The HF diet contained up to 60 % of calories from fat and was rich in stearic acid. Blood samples were collected at 0, 2, 4 and 6 weeks after the feeding. Adipose and liver tissues were collected at the 6(th) week after feeding. We performed analysis of histological findings and fatty acid composition in serum and tissues. RESULTS: Body weights of the cats significantly increased in the HF group. The increased activities of hepatic enzymes and the accumulation of lipid droplets were found in hepatocytes in the HF group at the 6(th) week after feeding. In this study, the stearic acid (C18:0)-rich HF diet contained less oleic acid (C18:1n-9) and more linoleic acid (C18:2n-6) than the control. However, the composition of oleic acid in the liver was higher, and those of stearic acid and linoleic acid were lower in the HF group at the 6(th) week after feeding. The higher oleic acid:stearic acid ratio suggests an increase in the conversion from saturated FA to mono-unsaturated FAs, which may reflect the hepatic storage of FAs as a relatively harmless form. CONCLUSION: The stearic acid-rich HF diet increased hepatic lipid accumulation accompanied by the increased of hepatic oleic acid, increased serum oleic acid and activation of hepatic enzymes. These findings could be an important sign of early stages of dyslipidemia and hepatic damage. Also, the higher oleic acid:stearic acid ratio might be related to the increased activity of SCD-1, which suggests that the stearic acid-rich HF diet evoked hepatic lipogenesis in the feline liver.

"Double-trick" visual and chemical mimicry by the juvenile orchid mantis hymenopus coronatus used in predation of the oriental honeybee apis cerana.

It has long been hypothesized that the flower-like appearance of the juvenile orchid mantis is used as visual camouflage to capture flower-visiting insects, although it is doubtful whether such morphological resemblance alone could increase their success in hunting. We confirmed that juvenile female orchid mantes often succeed in capturing oriental honeybees, while adult females often fail. Since most of the honeybees approached the juveniles from the front, we hypothesized that juvenile orchid mantes might attract honeybees by emitting some volatile chemical cues. Gas chromatography-mass spectrometry analyses revealed that the mantes' mandibular adducts contained 3-hydroxyoctanoic acid (3HOA) and 10-hydroxy-(E)-2-decenoic acid (10HDA), both of which are also features of the pheromone communication of the oriental honeybee. We also successfully detected 3HOA emitted in the head space air only at the time when the juvenile mantes were attempting to capture their prey. Field bioassay showed that the Oriental Honeybee predominantly preferred to visit dummies impregnated with a mixture of the appropriate amounts and ratios of 3HOA and 10HDA. We therefore conclude that the juvenile mantes utilize these as allelochemicals to trick and attract oriental honeybees.

Comparison of plasma lipoprotein profiles and malondialdehyde between hyperlipidemia dogs with/without treatment.

BACKGROUND: The aim of this study is to compare metabolic parameters, malondialdehyde as a lipid oxidation marker, and lipid profiles between dogs with untreated hyperlipidemia and hyperlipidemia with treatment, in order to examine the usefulness of malondialdehyde and lipid profiles as diagnostic parameters at early stages of hyperlipidemia. RESULTS: Dog samples were collected from four different veterinary clinics across Japan from March to June 2013. They were separated into three groups: control, untreated hyperlipidemia based on temporally screening, and hyperlipidemia with current anti-hyperlipidemic (statins and fibrates) treatment. Triglyceride levels of untreated hyperlipidemia dogs were significantly higher than those of control dogs. ALT levels of hyperlipidemic dogs with treatment were the highest among three groups. VLDL and LDL of both cholesterol and triglyceride of untreated hyperlipidemia dogs were the highest among three groups. HDL1 levels in triglyceride of hyperlipidemia dogs with treatment were significantly higher than those of control and untreated hyperlipidemia dog. Malondialdehyde concentrations of untreated hyperlipidemia dogs were significantly higher than those of control and hyperlipidemic dogs with treatment. CONCLUSIONS: In this study, dogs with untreated hyperlipidemia clearly showed abnormal lipid status, whereas hyperlipidemic dogs under anti-hyperlipidemia treatment showed more normal lipid status suggesting the effectiveness of the therapy. Anti-hyperlipidemics (statins and fibrates) for dogs are also effective in relieving elevated levels of lipids and lipid oxidation. Plasma lipid (triglyceride and cholesterol) profiles and malondialdehyde are useful diagnostic tools for identifying early stages of untreatment hyperlipidemia in dogs.

Age effects on plasma cholesterol and triglyceride profiles and metabolite concentrations in dogs.

BACKGROUND: In dogs, occurrence of lipid metabolism disorders such as obesity and diabetes mellitus has increased markedly in recent years. Hyperlipidemia has been regarded as a common characteristic for obese animals and hyperlipidemic condition may be associated with inflammation, oxidative stress and lipid composition changes. In this study, we investigated the changes in plasma cholesterol and triglyceride (TG) profiles and metabolite concentrations in 24 dogs (young group: 0-7 years old, n = 12, aged group: 8-13 years old, n = 12). RESULTS: Plasma adiponectin (ADN) concentrations were significantly lower in aged dogs than those in young dogs (mean ± SD, 17.2 ± 10.0 µg mL-1 vs 29.3 ± 12.5 µg mL-1, respectively; P <0.05). Although there were no significant differences statistically, aged dogs showed significantly higher plasma alpha1- acid glycoprotein (alpah1-AG) levels compared to those in young dogs. Plasma cholesterol lipoprotein and TG lipoprotein were divided into four fractions by biphasic agarose gel electrophoresis technique. The levels of the third TG-lipoprotein fraction from the positive pole (TG Fraction 3) were significantly higher in aged dogs than in young dogs (mean ± SD, 143.0 ± 109.3 mg dL-1 vs 55.2 ± 31.3 mg dL-1, respectively; P <0.05). On the correlation coefficient analysis by Peason's method, moderate positive correlations were seen between the age and TG (r = 0.446, P = 0.029), TG Fraction 3 (r = 0.516, P = 0.010), malondialdehyde (r = 0.146, P = 0.043), alpha-1 AG (r = 0.448, P = 0.028) levels, respectively. Moderate negative correlations were seen the age and total cholesterol (TC) Fraction 2 (r = -0.446, P = 0.029), glucose (r = -0.637, P = 0.001), ADN (r = -0.408, P = 0.048), respectively. CONCLUSIONS: Present data suggest biochemical characteristics of lipid metabolism disorder may be affected by aging in dogs.

Change in mRNA expression of sirtuin 1 and sirtuin 3 in cats fed on high fat diet.

BACKGROUND: Mammalian sirtuins are homologs to the yeast silent information regulator 2 (Sir2), which is an NAD-dependent deacetylase. Sirtuins are comprised of 7 proteins, and each has different target proteins. Sirtuin 1 (SIRT1) plays important roles in maintaining metabolic functions and immune responses, and SIRT3 protects cells from oxidative stress-induced cell death. Both SIRT1 and SIRT3 are regulated by metabolic status and aging. Hence, SIRT1 and SIRT3 have been researched in metabolic diseases, such as type 2 diabetes mellitus (DM), fatty liver, and heart diseases. Although these diseases have been increasing, there is little information about relation between the diseases and SIRT1 and SIRT3 in cats. Therefore we cloned SIRT1 and SIRT3 cDNA, examined mRNA expression in cat tissues, and investigated the changes in SIRT1 and SIRT3 mRNA expression in peripheral blood leukocyte of cats fed on HFD for 6 weeks. RESULTS: Cat SIRT1 and SIRT3 contained a catalytic core region and showed high sequence homology with other vertebrate SIRT1 (>61.3%) and SIRT3 (>65.9%) amino acids. Real-time polymerase chain reaction analyses revealed that high expression levels were observed in the liver and skeletal muscle for SIRT1 and in the heart for SIRT3 in cats. In addition, both cat SIRT1 and SIRT3 expression levels in the pancreas were different between individuals. Cat SIRT1 mRNA expression in peripheral blood leukocytes was significantly elevated in obese cats fed on HFD (P < 0.05). CONCLUSIONS: Cat SIRT1 and SIRT3 genes are highly conserved among vertebrates, and HFD feeding may be related to SIRT1 mRNA expression mechanisms in cat peripheral blood leukocytes.

Mechanism of insulin production in canine bone marrow derived mesenchymal stem cells.

Insulin is a critical hormone in the regulation of blood glucose levels and is produced exclusively by pancreatic islet beta-cells. Insulin deficiency due to reduced pancreatic islet beta-cell number underlies the progression of diabetes mellitus, prompting efforts to develop beta-cell replacement therapies. However, precise information on beta-cell replacement and differentiation in canines is limited. In this study, we established insulin-producing cells from bone marrow derived mesenchymal stem cells transiently expressing canine pancreatic and duodenal homeobox 1 (Pdx1), beta cell transactivator 2 (Beta2) and V-maf avian musculoaponeurotic fibrosarcoma oncogene homolog A (Mafa) using a gene transfer technique. Real-time PCR analysis revealed an increase in insulin mRNA expression of transfected cells. And ELISA revealed that insulin protein expressed was detected in cytoplasmic fraction. Insulin immunostaining analysis was performed and observed in cytoplasmic fraction. These results suggest that co-transfection of Pdx1, Beta2 and Mafa induce insulin production in canine BMSCs. Our findings provide a clue to basic research into the mechanisms underlying insulin production in the canines.

Comparison of bone marrow and adipose tissue-derived canine mesenchymal stem cells.

BACKGROUND: Bone marrow-derived mesenchymal stem cells (BM-MSCs) and adipose tissue-derived mesenchymal stem cells (AT-MSCs) are potential cellular sources of therapeutic stem cells. MSCs are a multipotent population of cells capable of differentiating into a number of mesodermal lineages. Treatment using MSCs appears to be a helpful approach for structural restoration in regenerative medicine. Correct identification of these cells is necessary, but there is inadequate information on the MSC profile of cell surface markers and mRNA expression in dogs. In this study, we performed molecular characterization of canine BM-MSCs and AT-MSCs using immunological and mRNA expression analysis. RESULTS: Samples were confirmed to be multipotent based on their osteogenic and adipogenic differentiation. And these cells were checked as stem cell, hematopoietic and embryonic stem cell (ESC) markers by flow cytometry. BM- and AT-MSCs showed high expression of CD29 and CD44, moderate expression of CD90, and were negative for CD34, CD45, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81. SSEA-1 was expressed at very low levels in AT-MSCs. Quantitative real-time PCR (qRT-PCR) revealed expression of Oct3/4, Sox2, and Nanog in BM- and AT-MSCs. There was no significant difference in expression of Oct3/4 and Sox2 between BM-MSCs and AT-MSCs. However, Nanog expression was 2.5-fold higher in AT-MSCs than in BM-MSCs. Using immunocytochemical analysis, Oct3/4 and Sox2 proteins were observed in BM- and AT-MSCs. CONCLUSION: Our results provide fundamental information to enable for more reproducible and reliable quality control in the identification of canine BM-MSCs and AT-MSCs by protein and mRNA expression analysis.

Primary pulmonary Ewing's sarcoma: report of a case.

The Ewing's sarcoma family of tumors has been reported to originate in a variety of sites, most commonly in the extremities. We herein describe a rare case of primary pulmonary Ewing's sarcoma in a patient with a family history of sarcoma. The patient was a 42-year-old male, who presented with hemoptysis. Chest radiographs revealed a pulmonary mass in the right lower lobe. Clinical and radiological examinations (computed tomography and positron emission tomography) revealed that the lesion was a primary lesion. The lesion was resected by right lower lobectomy. The tumor was located in the pulmonary parenchyma, and there was no evidence of an extrapulmonary involvement by the tumor. Histologically, the tumor was composed of uniform cells with round nuclei and scant cytoplasm which were arranged in cohesive lobules with rare pseudorosette formation. Immunohistochemically, the tumor cells were positive for CD99, and negative for epithelial markers, neuroendocrine markers, myogenic markers and lymphoma markers. This diagnosis was further supported by the cytogenic and reverse transcriptase-polymerase chain reaction findings of EWS/FLI-1 fusion transcripts. This demonstrated the presence of a very rare primary pulmonary Ewing's sarcoma. The patient was treated with chemotherapy after the operation because Ewing's sarcoma is an aggressive neoplasm. The patient has had no recurrent disease for 6 months after the operation.

Long-Term Follow-up With Multispecialty Management of a Giant Lymphangioma of an Infant Tongue Contributed to Reduced Complications of the Disease: A Case Report of a 21-Year Follow-up.

Background. Lymphangiomas are benign tumors of abnormal lymphatic tissue. Approximately 6% of all lymphangiomas occur on the tongue. A lymphangioma of the tongue may present as a localized or a diffused growth, which may enlarge to cause macroglossia, impaired speech, and difficulty in mastication. This article reports a 21-year follow-up of a male infant who presented with a giant tongue lymphangioma. This long-term follow-up with multidisciplinary management including partial glossectomy, sclerotherapy, and orthodontic treatment to diminish complications of the disease in adulthood.

Two chicken neuromedin U receptors: characterization of primary structure, biological activity and tissue distribution.

Neuromedin U (NMU) is a bioactive peptide that is involved in a variety of physiological functions. Two of its receptors, NMUR1 and NMUR2, have been identified and characterized in mammals. In this study, we performed cDNA cloning of chicken NMUR1 and NMUR2, and characterized their primary structure, biological activity, and expression patterns in chicken tissues. The chicken NMUR1 and NMUR2 cDNAs encoded 438 and 395 amino acid sequences, respectively. Chicken NMUR1 showed 54.8%-56.5% sequence identity with human, rat, and mouse NMUR1, and NMUR2 shared 67.3%-70.1% sequence identity with mammalian orthologs. Both chicken receptors have typical characteristics of G-protein-coupled receptors with seven transmembrane domains and the D/ERY motif. An increase in intracellular Ca(2+) mobilization was observed in HEK293 cells transfected with chicken NMUR1 or NMUR2 cDNA and treated with chicken or rat NMU. Real-time PCR analysis revealed that NMUR1 mRNA was preferentially expressed in the intestinal tissues such as the duodenum, jejunum, ileum, cecum, and colon/rectum, and brain regions such as the midbrain and optic lobe, and the ovary in adult hens. NMUR2 mRNA was exclusively expressed in the brain regions such as the cerebrum and midbrain. These results indicate that NMUR1 and NMUR2 mRNAs, which encode functional receptor proteins, are expressed in chicken tissues with different distribution patterns.

Molecular characterization of structure and tissue distribution of chicken neurotensin receptor.

Neurotensin, a tridecapeptide, is distributed in a wide range of tissues and exhibits multiple functions through its receptors. Hitherto molecular characterization of the neurotensin receptor has been reported in mammalian, amphibian, and fish species but not in avian species. In this study, we cloned the cDNA encoding chicken neurotensin receptor from the duodenum and characterized its primary structure, biological activity and distribution in the gastrointestinal tract. The cDNA encoded a protein consisting of 399 amino acids that had significant overall sequence homology to other vertebrate neurotensin receptor 1 with higher extent in the seven transmembrane domains. Chicken neurotensin increased intracellular Ca(2+) concentrations in human embryonic kidney 293 cells transiently expressing the chicken neurotensin receptor 1. Real-time PCR analysis showed that chicken neurotensin receptor 1 mRNA is expressed throughout the gastrointestinal tract with markedly higher level in the colon/rectum. These results indicate that the chicken neurotensin receptor 1 is involved in gastrointestinal functions through an intracellular signaling pathway accompanied by an increase in Ca(2+) levels.

Molecular characterization of free fatty acid receptors FFAR2 and FFAR3 in the domestic cat.

G protein-coupled receptors 41 and 43 were identified and characterized as free fatty acid receptors (FFAR) 3 and 2, respectively. FFAR2 and FFAR3 mediate short-chain fatty acids (SCFAs) as signalling molecules. The present study aimed to give molecular characterization of FFAR2 and FFAR3 in the domestic cat. High homology with that in other mammals was revealed by cDNA cloning of cat FFAR2 FFAR3. We analyzed the tissue distribution of cat FFAR2 and FFAR3 mRNA using quantitative polymerase chain reaction. The inhibition of intracellular cAMP concentrations was observed in cells transfected with cat FFAR2 or FFAR3 and treated with SCFAs. The activation of nuclear factor of activated T cells-luciferase reporter was only observed in cat FFAR2 transfected cells but not in FFAR3. Split luciferase assay (NanoLuc Binary Technology; NanoBiT) for FFAR2 or FFAR3 and Arrestin-3/ß-arrestin-2 revealed acetate-/propionate-induced recruitment to cat FFAR2 or FFAR3 in CHO-K1 cells. Our results indicate that FFAR2 and FFAR3 are functional receptor proteins that are expressed in cat tissues and show differential distribution patterns.