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1.
Respirol Case Rep ; 11(11): e01232, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37840601

ABSTRACT

We share our experiences of instructing three patients with severe upper limb dysfunction on how to self-adjust CPAP/NPPV masks. In Case 1, we simplified the procedure by suturing a part of the headband as the left forearm was amputated. In Case 2, the patient had congenitally short limbs with short stature; thus, we provided an additional belt to the headband to maintain the headband's configuration while wearing the mask. In Case 3, the patient had left hemiplegia due to stroke and, repetitive coaching was conducted during the recovery phase rehabilitation program. Difficulties with self-adjusting NPPV/CPAP masks can occur whenever there is limited hand mobility above the head, including upper limb dysfunction. Simplifying procedures and providing sufficient time for instruction could help achieve independence. There have been no previous reports describing similar training details. We believe that sharing this knowledge will be helpful to both patients and healthcare professionals.

2.
PLoS One ; 18(4): e0284353, 2023.
Article in English | MEDLINE | ID: mdl-37075046

ABSTRACT

Candida albicans can cause two major types of infections: superficial infection and systemic candidiasis. C. albicans infects diverse host niches, owing to a wide range of virulence factors and attributes, such as morphological transitions and phenotypic switching. C. albicans uses glycolysis, followed by alcoholic fermentation or mitochondrial respiration to rapidly generate ATP under aerobic conditions. In this study, we quantified the mRNA expression of several glycolysis-related enzymes associated with the initial phase of environmental changes using two strains: a type strain, NBRC 1385, and a strain from a patient with auto-brewery syndrome, LSEM 550. Additionally, we analyzed the regulation of a rate-limiting enzyme in glycolysis, phosphofructokinase 1 (PFK1). Our results showed that the mRNA expression of enzymes in the middle and last stages of glycolysis and alcoholic fermentation increased, and that of mitochondrial respiration enzymes decreased under short-term anaerobic conditions. Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) administration showed similar results under anaerobic conditions. Moreover, PFK1 maintained its regulatory effect under different conditions; no significant change was observed in its mRNA expression. Our results suggest that C. albicans obtains energy via carbohydrate catabolism in the early phase of environmental change and survives in various parts of the host.


Subject(s)
Candida albicans , Candidiasis , Humans , Anaerobiosis , Glycolysis , RNA, Messenger/metabolism
3.
J Vet Med Sci ; 85(3): 334-339, 2023 Mar 01.
Article in English | MEDLINE | ID: mdl-36725029

ABSTRACT

Previous studies on dolphin electrocardiograms have shown that they are mainly composed of increased negative waves, similar to ungulates. The electrocardiogram waveform was determined by the distribution of the Purkinje fibers. Based on the waveform of the dolphin electrocardiogram, Hamlin predicted that the Purkinje fibers would be distributed within the ventricular muscle, as in ungulates. The purpose of this study was to confirm the histological distribution of Purkinje fibers in dolphins. In the present study, bottlenose dolphin hearts were observed both grossly and histologically, and the effects of Purkinje fiber distribution and cardiac morphology on electrocardiogram waveforms were examined. This study showed that the Purkinje fibers of dolphins run just below the endocardium, as in humans, dogs, and cats, whose electrocardiograms mainly show positive waves. When the cardiac morphology of dolphins was observed carefully, the right ventricle was found to be extremely dilated compared to that of terrestrial mammals. In human recreational divers, right ventricular dilatation is induced by diving. We hypothesized that the dolphin's heart is in a state similar to that of the right heart dilatation in terrestrial animals. The dolphin electrocardiogram waveform was considered to be due to right axis deviation. Based on the above, we concluded that the dolphin electrocardiogram waveform was due to its ability to live in water. We found that the dolphins are genetically related to ungulates, particularly the hippopotamus, but that their hearts have evolved differently.


Subject(s)
Bottle-Nosed Dolphin , Animals , Humans , Dogs , Bottle-Nosed Dolphin/physiology , Thorax , Mammals , Electrocardiography , Heart Ventricles
4.
Vet Immunol Immunopathol ; 241: 110336, 2021 Nov.
Article in English | MEDLINE | ID: mdl-34649042

ABSTRACT

Neutrophils are essential for innate immunity as the first line of defence. Neutrophils act as phagocytic white blood cells to kill bacteria and other microorganisms. A strong respiratory burst of neutrophils, dependent on reactive oxygen species, is produced during phagocytosis. Platelet-activating factor (PAF) is a signalling molecule with several prominent roles in tissue injury, inflammation, and platelet aggregation. However, the detailed mechanisms and intracellular signalling pathways involved in PAF-mediated neutrophil activation remain unclear. Here, we investigated the effect of PAF on changes in calcium concentration ([Ca2+]i) and oxygen radical (O2-) generation in activating canine neutrophils. We further evaluated these effects of PAF with inhibition of G protein-coupled receptors using the specific inhibitor suramin. Blood samples were collected from a total of five dogs and neutrophils were isolated. PAF stimulation of canine neutrophils caused an increase in [Ca2+]i as well as the generation of O2-, and the PAF receptor was sensitive to suramin. The results suggested that PAF stimulation of canine neutrophils may cause Ca2+ influx from the endoplasmic reticulum into the cytoplasm (as the first wave) and then trigger store-operated Ca2+ entry (as the second wave), which is an important intracellular signal transduction pathway for neutrophil activation. Furthermore, O2- generation by PAF stimulation may depend on the intracellular signalling pathway, with increasing inositol trisphosphate levels and [Ca2+]i via G protein-coupled receptors. The finding that PAF-activating platelet aggregation is involved in canine neutrophil activation suggests a close relationship between haemostasis and neutrophil activation in dogs, offering new insight into the response to infection.


Subject(s)
Neutrophils , Platelet Activating Factor , Animals , Calcium , Dogs , Neutrophils/cytology , Platelet Activating Factor/pharmacology , Reactive Oxygen Species , Receptors, G-Protein-Coupled , Signal Transduction , Suramin/pharmacology
5.
Arch Oral Biol ; 132: 105276, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34634536

ABSTRACT

OBJECTIVE: The physiological expression of cell division cycle 42 (cdc42) in major salivary glands, and paracellular transport of fluorescein isothiocyanate-dextran (FITC-dextran) in SMIE cells, which regulate cdc42 expression, was investigated to clarify the involvement of cdc42 in salivary production. DESIGN: The physiological expression of cdc42 in the rat submandibular gland, parotid gland, sublingual gland, and SMIE cells was detected using SDS-PAGE and western blotting. The paracellular transport of FITC-dextran in transwells was compared in transfected SMIE cells, exhibiting up- or downregulated cdc42 expression. RESULTS: Cdc42 was expressed in all major salivary glands and SMIE cells. SMIE cells transfected with the cdc42 plasmid had an increase efflux. In addition, SMIE cells transfected with the cdc42 siRNA showed decreased efflux. CONCLUSION: We suggest that cdc42 enhances paracellular transport in salivary glands without any morphological changes, including cell-cell adhesion.


Subject(s)
Sublingual Gland , Submandibular Gland , cdc42 GTP-Binding Protein , Animals , Blotting, Western , Parotid Gland , Rats , Salivary Glands
6.
Arch Oral Biol ; 129: 105209, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34274846

ABSTRACT

OBJECTIVE: Secretory immunoglobulin A (sIgA) is important for mucosal immunity due to the inhibition of pathogen infection. The submandibular gland is known to secrete more sIgA than the parotid and sublingual glands. In this study, we focused on the relationship between the secretion of accumulated intracellular sIgA and ß-adrenergic receptor stimulation, and clarified the autonomic regulatory mechanism of sIgA secretion in submandibular gland cells using dispersed gland cells. DESIGN: Sprague-Dawley rats (male, 6 weeks old, 200-250 g) were euthanized and their submandibular glands were removed. Dispersed submandibular gland cells placed in Krebs-Ringer-Bicarbonate solution were stimulated by autonomic nerve agonists. The concentration of secreted sIgA was measured using a rat IgA ELISA kit. The results were analysed using ANOVA and Tukey's test. RESULTS: Cells stimulated with the non-selective ß-adrenoreceptor agonist, isoprenaline, secreted significantly more sIgA compared with the unstimulated control. The ß2-adrenoreceptor agonist, fenoterol, caused significantly more sIgA secretion than the control, and more sIgA secretion than the ß1-adrenoreceptor agonist, xamoterol. sIgA secretion by isoprenaline stimulation was dose dependent. Inhibition of the ß receptor by propranolol completely blocked sIgA secretion following isoprenaline stimulation. CONCLUSION: Stimulation of ß receptors could result in more secretion of intracellularly accumulated sIgA compared with stimulation of other autonomic receptors in the autonomic modulation of mucosal immunity.


Subject(s)
Immunoglobulin A, Secretory , Submandibular Gland , Adrenergic beta-Agonists/pharmacology , Animals , Isoproterenol/pharmacology , Male , Parotid Gland , Rats , Rats, Sprague-Dawley , Receptors, Adrenergic, beta , Saliva
7.
J Vet Med Sci ; 81(10): 1515-1521, 2019 Oct 24.
Article in English | MEDLINE | ID: mdl-31474665

ABSTRACT

In healthy individuals, plasma glucose levels are maintained within a normal range. During fasting, endogenous glucose is released either through glycogenolysis or gluconeogenesis. Gluconeogenesis involves the formation of glucose-6-phosphate from a variety of precursors followed by its subsequent hydrolysis to glucose. Gluconeogenesis occurs in the liver and the kidney. In order to compare gluconeogenesis in canine liver and kidney, the activity and expression of the rate limiting enzymes that catalyze the fructose-6-phosphate and fructose 1,6-bisphosphate steps, namely, phosphofructokinase-1 (PFK-1) (glycolysis) and fructose bisphosphatase-1 (FBP-1) (gluconeogenesis), were examined. Healthy male and female beagle dogs aged 1-2 years were euthanized humanely, and samples of their liver and kidney were obtained for analysis. The levels of PFK-1 and FBP-1 in canine liver and kidney were assessed by enzymatic assays, Western blotting, and RT-qPCR. Enzyme assays showed that, in dogs, the kidney had higher specific activity of PFK-1 and FBP-1 than the liver. Western blotting and RT-qPCR data demonstrated that of the three different subunits (PFK-M, PFK-L, and PFK-P) the PFK-1 in canine liver mainly comprised PFK-L, whereas the PFK-1 in the canine kidney comprised all three subunits. As a result of these differences in the subunit composition of PFK-1, glucose metabolism might be regulated differently in the liver and kidney.


Subject(s)
Fructose-Bisphosphatase/metabolism , Kidney/enzymology , Liver/enzymology , Phosphofructokinase-1/metabolism , Animals , Dogs , Female , Fructose-Bisphosphatase/chemistry , Gluconeogenesis , Glycolysis , Male , Phosphofructokinase-1/chemistry
8.
J Vet Med Sci ; 81(5): 712-716, 2019 May 31.
Article in English | MEDLINE | ID: mdl-30918224

ABSTRACT

Phosphofructokinase-1 (EC:2.7.1.11, PFK-1) catalyzes the phosphorylation of fructose 6-phosphate to fructose 1,6-bisphosphate using adenosine triphosphate and is a key regulatory enzyme of glycolysis. Mammalian PFK-1 isozymes are composed of three kinds of subunits (PFK-M, -L, and -P), with different properties. It has been suggested that the proportion of PFK-1 subunits in different organs is based on the organ energy metabolism. In this study, we analyzed the activity and subunit composition of canine PFK-1. We found that, in dogs, the skeletal muscle only has PFK-M, the liver mainly has PFK-L, and the brain expresses all of them. The knowledge of the composition of PFK-1 could provide useful information for determination of the differences in glycolysis in various organs of dogs.


Subject(s)
Dogs/metabolism , Isoenzymes/metabolism , Phosphofructokinase-1/metabolism , Animals , Brain/enzymology , Female , Liver/enzymology , Male , Muscle, Skeletal/enzymology , Phosphofructokinase-1/chemistry , Tissue Distribution
9.
J Vet Med Sci ; 81(3): 369-372, 2019 Mar 14.
Article in English | MEDLINE | ID: mdl-30674739

ABSTRACT

This study was undertaken to establish a method for measuring mRNA expression by using real-time RT-PCR in the diagnosis of canine meningiomas. When performing real-time RT-PCR, it is essential to include appropriate control tissues and to select appropriate housekeeping genes as an internal standard. Based on the results of our study, RPS18 constitutes a suitable internal standard for the comparison of mRNA expression between normal meninges and meningiomas. The results showed increased mRNA expression of VEGFA and EGFR; however, mRNA expression of KDR was reduced. Measuring mRNA expression by using real-time RT-PCR with appropriate control tissues and internal standards can provide useful information to understanding the pathogenesis of canine meningiomas, which corresponds with immunohistochemical findings.


Subject(s)
Meningeal Neoplasms/veterinary , Meningioma/veterinary , RNA, Messenger/biosynthesis , RNA, Neoplasm/biosynthesis , Animals , Dogs , Female , Genes, Neoplasm , Male , Meningeal Neoplasms/genetics , Meningioma/genetics , Neoplasm Grading/veterinary , Real-Time Polymerase Chain Reaction/veterinary
10.
Sci Rep ; 7: 39914, 2017 01 05.
Article in English | MEDLINE | ID: mdl-28054591

ABSTRACT

The proinflammatory cytokine interleukin 1ß (IL-1ß) induces prostaglandin E2 (PGE2) production via upregulation of cyclooxygenase-2 (COX-2) expression in synovial fibroblasts. This effect of IL-1ß is involved in osteoarthritis. We investigated MAPK signaling pathways in IL-1ß-induced COX-2 expression in feline synovial fibroblasts. In the presence of MAPK inhibitors, IL-1ß-induced COX-2 expression and PGE2 release were both attenuated. IL-1ß induced the phosphorylation of p38, JNK, MEK, and ERK1/2. A JNK inhibitor prevented not only JNK phosphorylation but also MEK and ERK1/2 phosphorylation in IL-1ß-stimulated cells, but MEK and ERK1/2 inhibitors had no effect on JNK phosphorylation. A p38 inhibitor prevented p38 phosphorylation, but had no effect on MEK, ERK1/2, and JNK phosphorylation. MEK, ERK1/2, and JNK inhibitors had no effect on p38 phosphorylation. We also observed that in IL-1ß-treated cells, phosphorylated MEK, ERK1/2, and JNK were co-precipitated with anti-phospho-MEK, ERK1/2, and JNK antibodies. The silencing of JNK1 in siRNA-transfected fibroblasts prevented IL-1ß to induce phosphorylation of MEK and ERK1/2 and COX-2 mRNA expression. These observations suggest that JNK1 phosphorylation is necessary for the activation of the MEK/ERK1/2 pathway and the subsequent COX-2 expression for PGE2 release, and p38 independently contributes to the IL-1ß effect in synovial fibroblasts.


Subject(s)
Cyclooxygenase 2/metabolism , Extracellular Signal-Regulated MAP Kinases/metabolism , Fibroblasts/physiology , Interleukin-1beta/metabolism , MAP Kinase Kinase Kinases/metabolism , Mitogen-Activated Protein Kinase 8/metabolism , Signal Transduction , Animals , Cats , Cells, Cultured , Dinoprostone/metabolism
11.
PLoS One ; 11(11): e0166707, 2016.
Article in English | MEDLINE | ID: mdl-27851800

ABSTRACT

Acute kidney injury (AKI) is characterized by a sudden loss of renal function. Early recognition of AKI, especially in critically ill patients, is essential for adequate therapy. Currently, neutrophil gelatinase-associated lipocalin (NGAL) is considered to be an effective biomarker of AKI; however, the regulation of its expression and function in renal tubular cells remains unclear. In this study, we investigated the regulation of the expression and function of NGAL in IL-1ß-treated Madin-Darby canine kidney (MDCK) cells as a model of renal tubular cells. IL-1ß induced a disturbance in the localization of E-cadherin and zonaoccludin-1 (ZO-1). The transepithelial electrical resistance (TER) also decreased 5 days after IL-1ß treatment. IL-1ß induced NGAL mRNA expression and protein secretion in a time- and dose-dependent manner, which occurred faster than the decrease in TER. In the presence of ERK1/2 and p38 inhibitors, IL-1ß-induced NGAL mRNA expression and protein secretion were significantly attenuated. In the presence of recombinant NGAL, IL-1ß-induced disturbance in the localization of E-cadherin and ZO-1 was attenuated, and the decrease in TER was partially maintained. These results suggest that NGAL can be used as a biomarker for AKI and that it functions as a protector from AKI.


Subject(s)
Interleukin-1beta/metabolism , Kidney Tubules/cytology , Kidney Tubules/metabolism , Lipocalin-2/metabolism , Adherens Junctions/drug effects , Adherens Junctions/metabolism , Animals , Cadherins/metabolism , Dogs , Electric Impedance , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors , Extracellular Signal-Regulated MAP Kinases/metabolism , Kidney Tubules/drug effects , Lipocalin-2/genetics , Madin Darby Canine Kidney Cells , Protein Kinase Inhibitors/pharmacology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Recombinant Proteins/metabolism , Tight Junctions/drug effects , Tight Junctions/metabolism , Zonula Occludens-1 Protein/metabolism , p38 Mitogen-Activated Protein Kinases/metabolism
12.
Vet Immunol Immunopathol ; 168(3-4): 223-32, 2015 Dec 15.
Article in English | MEDLINE | ID: mdl-26549149

ABSTRACT

The proinflammatory cytokine interleukin-1ß (IL-1ß) induced cyclooxygenases-2 (COX-2) mRNA expression and lipid mediator prostaglandin E2 release and in a time- and dose-dependent manner in canine dermal fibroblasts. The MEK inhibitor U0126 and the ERK inhibitor FR180204 clearly inhibited IL-1ß-induced prostaglandin E2 release and COX-2 mRNA expression. IL-1ß enhanced ERK1/2 phosphorylation, which was attenuated by inhibitors of MEK and ERK. The NF-κB inhibitor BAY 11-7082 also suppressed IL-1ß-induced prostaglandin E2 release and COX-2 mRNA expression. Treatment of fibroblasts with IL-1ß led to the phosphorylation of p65 and degradation of IκBα occurred, indicating that IL-1ß treatment activated NF-κB. MEK and ERK1/2 inhibitors had no effect on the phosphorylation of p65 subunit induced by IL-1ß, whereas the NF-κB inhibitor completely blocked IL-1ß-induced phosphorylation of ERK1/2. We also observed that IκBα-knockdown enhanced the phosphorylation of p65 and ERK1/2. These findings suggest that stimulation of MEK/ERK signaling pathway by NF-κB activation regulates IL-1ß-induced COX-2 expression and subsequent prostaglandin E2 release in canine dermal fibroblasts.


Subject(s)
Cyclooxygenase 2/metabolism , Dogs , Fibroblasts/metabolism , Interleukin-1beta/metabolism , MAP Kinase Signaling System/physiology , NF-kappa B/metabolism , Animals , Cyclooxygenase 2/genetics , Dinoprostone/genetics , Dinoprostone/metabolism , Fibroblasts/drug effects , Gene Expression Regulation, Enzymologic , Interleukin-1beta/genetics , MAP Kinase Signaling System/genetics , Male , NF-kappa B/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Wound Healing/physiology
13.
PLoS One ; 10(11): e0141581, 2015.
Article in English | MEDLINE | ID: mdl-26523832

ABSTRACT

Bone marrow stromal cells (BMSCs) are considered as candidates for regenerative therapy and a useful model for studying neuronal differentiation. The role of basic fibroblast growth factor (bFGF) in neuronal differentiation has been previously studied; however, the signaling pathway involved in this process remains poorly understood. In this study, we investigated the signaling pathway in the bFGF-induced neuronal differentiation of canine BMSCs. bFGF induced the mRNA expression of the neuron marker, microtubule associated protein-2 (MAP2) and the neuron-like morphological change in canine BMSCs. In the presence of inhibitors of fibroblast growth factor receptors (FGFR), phosphatidylinositol 3-kinase (PI3K) and Akt, i.e., SU5402, LY294002, and MK2206, respectively, bFGF failed to induce the MAP2 mRNA expression and the neuron-like morphological change. bFGF induced Akt phosphorylation, but it was attenuated by the FGFR inhibitor SU5402 and the PI3K inhibitor LY294002. In canine BMSCs, expression of FGFR-1 and FGFR-2 was confirmed, but only FGFR-2 activation was detected by cross-linking and immunoprecipitation analysis. Small interfering RNA-mediated knockdown of FGFR-2 in canine BMSCs resulted in the attenuation of bFGF-induced Akt phosphorylation. These results suggest that the FGFR-2/PI3K/Akt signaling pathway is involved in the bFGF-induced neuronal differentiation of canine BMSCs.


Subject(s)
Fibroblast Growth Factor 2/pharmacology , Mesenchymal Stem Cells/drug effects , Neurons/cytology , Receptor, Fibroblast Growth Factor, Type 2/metabolism , Signal Transduction/drug effects , Animals , Cell Differentiation/drug effects , Cells, Cultured , Dogs , Gene Expression Regulation/drug effects , Male , Mesenchymal Stem Cells/cytology , Microtubule-Associated Proteins/genetics , Neurons/drug effects , Oncogene Protein v-akt/genetics , Oncogene Protein v-akt/metabolism , Phosphatidylinositol 3-Kinases/genetics , Phosphatidylinositol 3-Kinases/metabolism , Phosphorylation/drug effects , Receptor, Fibroblast Growth Factor, Type 2/genetics
14.
J Vet Med Sci ; 77(1): 27-35, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25284120

ABSTRACT

We investigated the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage- and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons containing recombinant human basic fibroblast growth factor (bFGF; 100 ng/ml). The viability of the bFGF-treated cells was assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial markers. Western blotting and immunocytochemical analysis for the neuronal markers were performed to evaluate the protein expression and localization. The Ca(2+) mobilization of the cells was evaluated using the Ca(2+) indicator Fluo3 to monitor Ca(2+) influx. To investigate the mechanism of bFGF-induced neuronal differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide 3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore, in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase in intracellular Ca(2+) levels. Each inhibitor significantly attenuated the bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells and may lead to the development of new cell-based treatments for neuronal diseases.


Subject(s)
Cell Differentiation/physiology , Fibroblast Growth Factor 2/pharmacology , Mesenchymal Stem Cells/physiology , Neurons/physiology , Animals , Cell Differentiation/drug effects , Cells, Cultured , Dogs , Gene Expression Regulation/drug effects , Gene Expression Regulation/physiology , Male , Mesenchymal Stem Cells/cytology , Phosphatidylinositol 3-Kinases/genetics , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/genetics , Proto-Oncogene Proteins c-akt/metabolism , Receptors, Fibroblast Growth Factor/genetics , Receptors, Fibroblast Growth Factor/metabolism
15.
Kekkaku ; 90(10): 671-5, 2015 Oct.
Article in Japanese | MEDLINE | ID: mdl-26821396

ABSTRACT

A 27-year-old man with a 4-month history of treatment for miliary tuberculosis at another hospital was admitted to our hospital for continued treatment. Computed tomography showed new lesions in the S8 area of the liver and spleen, despite resolution of chest radiographic findings. Because these new lesions were still present after 8 months of treatment, we performed laparoscopic drainage of the liver abscess. Purulent material drained from the lesion revealed positive polymerase chain reaction results for Mycobacterium tuberculosis, and identification of granuloma with infiltrating lymphocytes and plasma cells confirmed the diagnosis of tubercular liver abscess. Pathological changes in the spleen over the clinical course were also regarded as representing tubercular abscess. Postoperative course was good, and tuberculosis treatment ended after 12 months. Tubercular liver abscess subsequently showed prominent reduction, and the tubercular splenic abscess disappeared on abdominal ultrasonography. Tubercular hepatosplenic abscesses appearing during tubercular treatment are rare. We report this valuable case in which laparoscopic drainage of a liver abscess proved useful for diagnosis and treatment.


Subject(s)
Liver Abscess/etiology , Liver Abscess/therapy , Splenic Diseases/etiology , Splenic Diseases/therapy , Tuberculosis, Miliary/complications , Tuberculosis, Miliary/drug therapy , Tuberculosis/etiology , Tuberculosis/therapy , Abscess/diagnosis , Abscess/etiology , Abscess/therapy , Antitubercular Agents/administration & dosage , Drainage/methods , Humans , Laparoscopy , Liver Abscess/diagnosis , Male , Mycobacterium tuberculosis/isolation & purification , Splenic Diseases/diagnosis , Treatment Outcome , Tuberculosis/diagnosis , Tuberculosis/microbiology
16.
Metallomics ; 6(9): 1656-63, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24951051

ABSTRACT

ZnT-1 is a Cation Diffusion Facilitator (CDF) family protein, and is present throughout the phylogenetic tree from bacteria to humans. Since its original cloning in 1995, ZnT-1 has been considered to be the major Zn(2+) extruding transporter, based on its ability to protect cells against zinc toxicity. However, experimental evidence for ZnT-1 induced Zn(2+) extrusion was not convincing. In the present study, based on the 3D crystal structure of the ZnT-1 homologue, YiiP, that predicts a homodimer that utilizes the H(+) electrochemical gradient to facilitate Zn(2+) efflux, we demonstrate ZnT-1 dependent Zn(2+) efflux from HEK 293T cells using FluoZin-3 and Fura 2 by single cell microscope based fluorescent imaging. ZnT-1 facilitates zinc efflux in a sodium-independent, pH-driven and calcium-sensitive manner. Moreover, substitution of two amino acids in the putative zinc binding domain of ZnT-1 led to nullification of Zn(2+) efflux and rendered the mutated protein incapable of protecting cells against Zn(2+) toxicity. Our results demonstrate that ZnT-1 extrudes zinc from mammalian cells by functioning as a Zn(2+)/H(+) exchanger.


Subject(s)
Cation Transport Proteins/metabolism , Hydrogen/metabolism , Mammals/metabolism , Zinc/metabolism , Animals , Biological Transport/drug effects , Calcium/metabolism , Cell Membrane/drug effects , Cell Membrane/metabolism , Cytoprotection/drug effects , Electrochemistry , HEK293 Cells , Humans , Hydrogen-Ion Concentration , Mutant Proteins/metabolism , Sodium/metabolism , Zinc/toxicity
17.
Am J Vet Res ; 74(10): 1311-20, 2013 Oct.
Article in English | MEDLINE | ID: mdl-24066915

ABSTRACT

OBJECTIVE: To investigate the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into functional, mature neurons. SAMPLE: Bone marrow from 6 adult dogs. PROCEDURES: BMSCs were isolated from bone marrow and chemically induced to develop into neurons. The morphology of the BMSCs during neuronal induction was monitored, and immunocytochemical analyses for neuron markers were performed after the induction. Real-time PCR methods were used to evaluate the mRNA expression levels of markers for neural stem or progenitor cells, neurons, and ion channels, and western blotting was used to assess the expression of neuronal proteins before and after neuronal induction. The electrophysiological properties of the neuron-like cells induced from canine BMSCs were evaluated with fluorescent dye to monitor Ca(2)+ influx. RESULTS: Canine BMSCs developed a neuron-like morphology after neuronal induction. Immunocytochemical analysis revealed that these neuron-like cells were positive for neuron markers. After induction, the cells' mRNA expression levels of almost all neuron and ion channel markers increased, and the protein expression levels of nestin and neurofilament-L increased significantly. However, the neuron-like cells derived from canine BMSCs did not have the Ca(2)+ influx characteristic of spiking neurons. CONCLUSIONS AND CLINICAL RELEVANCE: Although canine BMSCs had neuron-like morphological and biochemical properties after induction, they did not develop the electrophysiological characteristics of neurons. Thus, these results have suggested that canine BMSCs could have the capacity to differentiate into a neuronal lineage, but the differentiation protocol used may have been insufficient to induce development into functional neurons.


Subject(s)
Cell Differentiation/physiology , Mesenchymal Stem Cells/metabolism , Neural Stem Cells/metabolism , RNA, Messenger/metabolism , Analysis of Variance , Animals , Blotting, Western/veterinary , Calcium Channels/metabolism , Dogs , Immunohistochemistry/veterinary , Mesenchymal Stem Cells/physiology , Nestin/metabolism , Neural Stem Cells/cytology , Neurofilament Proteins/metabolism , Real-Time Polymerase Chain Reaction/veterinary , Reverse Transcriptase Polymerase Chain Reaction/veterinary
18.
Pflugers Arch ; 465(2): 271-81, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23179381

ABSTRACT

Non-obese diabetic (NOD) mice have been used as a model for dry mouth. NOD mice lacking the gene encoding E2f1, a transcription factor, develop hyposalivation more rapidly progressively than control NOD mice. However, the model mice are associated with an underlying disease such as diabetes. We have now established E2f1-deficient NOD/severe combined immunodeficiency disease (NOD/SCID.E2f1(-/-)) mice to avoid the development of diabetes (Matsui-Inohara et al., Exp Biol Med (Maywood) 234(12):1525-1536, 2009). In this study, we investigated the pathophysiological features of dry mouth using NOD/SCID.E2f1(-/-) mice. In NOD/SCID.E2f1(-/-) mice, the volume of secreted saliva stimulated with pilocarpine is about one third that of control NOD/SCID mice. In behavioral analysis, NOD/SCID.E2f1(-/-) mice drank plenty of water when they ate dry food, and the frequency and time of water intake were almost double compared with control NOD/SCID mice. Histological analysis of submandibular glands with hematoxylin-eosin stain revealed that NOD/SCID.E2f1(-/-) mice have more ducts than NOD/SCID mice. In western blot analysis, the expression of aquaporin 5 (AQP5), a marker of acinar cells, in parotid and in submandibular glands of NOD/SCID.E2f1(-/-) mice was lower than in NOD/SCID mice. Immunohistochemical analysis of parotid and submandibular acini revealed that the localization of AQP5 in NOD/SCID.E2f1(-/-) mice differs from that in NOD/SCID mice; AQP5 was leaky and diffusively localized from the apical membrane to the cytosol in NOD/SCID.E2f1(-/-) mice. The ubiquitination of AQP5 was detected in submandibular glands of NOD/SCID.E2f1(-/-) mice. These findings suggest that the change of acinar/duct structure and the down-regulation of AQP5 in the salivary gland cause the pathogenesis of hyposalivation in NOD/SCID.E2f1(-/-) mice.


Subject(s)
Acinar Cells/metabolism , Aquaporin 5/metabolism , Down-Regulation , E2F1 Transcription Factor/genetics , Salivary Ducts/metabolism , Xerostomia/metabolism , Acinar Cells/pathology , Animals , Aquaporin 5/genetics , Cell Membrane/metabolism , Cytosol/metabolism , Drinking , Gene Expression , Mice , Mice, Inbred NOD , Mice, Knockout , Mice, SCID , Parotid Gland/metabolism , Parotid Gland/pathology , Pilocarpine/pharmacology , Salivary Ducts/pathology , Salivation/drug effects , Salivation/genetics , Submandibular Gland/metabolism , Submandibular Gland/pathology , Ubiquitination , Xerostomia/genetics , Xerostomia/physiopathology
19.
J Oral Sci ; 54(1): 23-32, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22466883

ABSTRACT

Polymeric immunoglobulin receptor (pIgR) plays an intrinsic role in protecting the intestinal tract from invading pathogens. In the present study, we observed a decrease in pIgR in colon lysate from mice with dextran sodium sulfate (DSS) colitis. A decrease in pIgR was detected in both mRNA and protein levels. Histologic examinations revealed marked destruction of intestinal epithelial cells (IECs), and only a small number of regenerating IECs expressed pIgR. These results suggest that the decrease in pIgR was due to the destruction of IECs. Because activation of toll-like receptor 3 slows the progression of DSS colitis, we injected polyriboinosinic: polyribocytidylic acid (poly I:C) intraperitoneally and observed the correlation between pIgR level and severity of DSS colitis. Poly I:C markedly decreased progression of DSS colitis, and pIgR levels significantly recovered. Furthermore, we found that expressions of IFN-γ and TNF-α were higher in DSS colitis. These results indicate that the decrease in pIgR was not compensated for by increased expression of these cytokines. In sum, our findings show that pIgR levels vary according to the severity of DSS colitis and that these changes might be useful as a biomarker of the severity of inflammatory bowel disease.


Subject(s)
Colitis/metabolism , Receptors, Polymeric Immunoglobulin/biosynthesis , Animals , Biomarkers , Colitis/chemically induced , Dextran Sulfate , Female , Interferon-gamma/biosynthesis , Interleukin-10/biosynthesis , Interleukin-4/biosynthesis , Intestinal Mucosa/cytology , Intestinal Mucosa/metabolism , Mice , Mice, Inbred C57BL , Poly I-C/pharmacology , Toll-Like Receptor 3/antagonists & inhibitors , Tumor Necrosis Factor-alpha/biosynthesis
20.
Kekkaku ; 86(2): 57-60, 2011 Feb.
Article in Japanese | MEDLINE | ID: mdl-21404651

ABSTRACT

A 49-year-old male who had been treated for pulmonary tuberculosis and tuberculous pleurisy in 2007 was referred to our hospital with the complaint of dyspnea on exertion in Nov. 2009. Chest X-ray showed increased pleural effusion compared with that remaining after the previous treatment of pleurisy in 2008. A chest CT revealed that fluid collection was surrounded by thickened pleura. Thoracocentesis was performed, and yellow milky liquid was obtained. The pleural effusion contained few cells. The triglyceride concentration was 83 mg/dl, and the cholesterol level was very high at 628 mg/dl. Based on these findings we diagnosed this case as chyliform pleural effusion. Both smear of acid-fast bacilli and PCR-TB test of the pleural effusion were positive, but culture was negative for mycobacterium, suggesting that this chyliform pleural effusion was produced by the former episode of tuberculous pleurisy, not by the recent reactivation of tuberculous pleurisy. The ADA concentration in the pleural effusion was high at 91.7 IU/l. No increase in the amount of pleural effusion was observed after thoracocentesis without any anti-tuberculosis therapy.


Subject(s)
Pleural Effusion/etiology , Tuberculosis, Pleural/complications , Chyle , Humans , Male , Middle Aged
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