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Biomedical subjects

T Kubo

Publications and source records attributed to T Kubo.

At least 235 records · Page 13Linked to original sources

Expression of cytoplasmic galectin-3 as a prognostic marker in tongue carcinoma.

Galectin-3 is a member of the beta-galactoside-binding mammalian lectin family with affinity to ABH group epitopes, cell surface and extracellular polylactosamine glycans. It has been shown to be involved in differentiation, morphogenesis, tumor progression, and metastasis. Here we questioned the possible involvement of galectin-3 in the neoplastic progression of the tongue epithelium and evaluated its prognostic value in tongue cancer patients. Galectin-3 expression was analyzed by the immunohistochemical method in 77 tongue specimens (54 squamous cell carcinomas and 23 specimens of distinct normal mucosa). Levels of nuclear expression of galectin-3 markedly decreased during the progression from normal to cancerous states (P < 0.0001), while cytoplasmic expression increased (P < 0.0001). Enhanced expression of galectin-3 in the cytoplasm was associated with a reduced disease-free survival of tongue cancer patients. Multivariate analysis identified enhanced expression of cytoplasmic galectin-3 as an independent predictor of disease recurrence (P = 0.0120). These results suggest that the observed translocation of galectin-3 from the nucleus to the cytoplasm during neoplastic progression may serve as a prognostic factor for tongue cancer patients.

Adult↗

Effects of hypergravity on morphology and osteopontin expression in the rat otolith organs.

Recovery from balance disorders after spaceflight is supposed to reflect the process of readaptation to normal gravitational environment (1G) from microgravity. Linear acceleration including gravity is perceived by otolith pathways, therefore, it could be assumed that possible plastic changes in any part of otolithic pathways from the level of the vestibular periphery to the central vestibular system might be responsible for adaptive mechanisms to an altered gravitational environment. In the present study, to elucidate a role of otoconia in adaptation to altered gravity, we examined the effects of hypergravity (2G) on morphology and synthesis of saccular and utricular otoconia in young adult rats. Morphology of otoconia was examined by scanning electron microscopy. Otoconial synthesis was assessed by mRNA expression of osteopontin, a matrix protein of otoconia, in otolithic maculae as a marker of otoconial genesis determined by a real-time quantitative PCR method. The present results showed that neither otoconial morphology nor otoconial synthesis was affected by up to one week exposure to hypergravity. These findings suggest that changes in neurotransmission at the synapses of the peripheral and/or central vestibular system rather than the changes in otoconial morphology and synthesis may be involved in adaptive mechanisms to an altered gravitational environment.

Animals↗

Soldier caste-specific gene expression in the mandibular glands of Hodotermopsis japonica (Isoptera: termopsidae).

Although "polymorphic castes" in social insects are well known as one of the most important phenomena of polyphenism, few studies of caste-specific gene expressions have been performed in social insects. To identify genes specifically expressed in the soldier caste of the Japanese damp-wood termite Hodotermopsis japonica, we employed the differential-display method using oligo(dT) and arbitrary primers, compared mRNA from the heads of mature soldiers and pseudergates (worker caste), and identified a clone (PCR product) 329 bp in length termed SOL1. Northern blot analysis showed that the SOL1 mRNA is about 1.0 kb in length and is expressed specifically in mature soldiers, but not in pseudergates, even in the presoldier induction by juvenile hormone analogue, suggesting that the product is specific for terminally differentiated soldiers. By using the method of 5'- and 3'-rapid amplification of cDNA ends, we isolated the full length of SOL1 cDNA, which contained an ORF with a putative signal peptide at the N terminus. The sequence showed no significant homology with any other known protein sequences. In situ hybridization analysis showed that SOL1 is expressed specifically in the mandibular glands. These results strongly suggest that the SOL1 gene encodes a secretory protein specifically synthesized in the mandibular glands of the soldiers. Histological observations revealed that the gland actually develops during the differentiation into the soldier caste.

Amino Acid Sequence↗

Expression of amylase and glucose oxidase in the hypopharyngeal gland with an age-dependent role change of the worker honeybee (Apis mellifera L.).

Worker honeybees change their behaviour from the role of nurse to that of forager with age. We have isolated cDNA clones for two honeybee (Apis mellifera L.) genes, encoding alpha-amylase and glucose oxidase homologues, that are expressed in the hypopharyngeal gland of forager bees. The predicted amino acid sequence of the putative Apis amylase showed 60.5% identity with Drosophila melanogaster alpha-amylase, whereas that of Apis glucose oxidase showed 23.8% identity with Aspergillus niger glucose oxidase. To determine whether the isolated cDNAs actually encode these enzymes, we purified amylase and glucose oxidase from homogenized forager-bee hypopharyngeal glands. We sequenced the N-terminal regions of the purified enzymes and found that they matched the corresponding cDNAs. mRNAs for both enzymes were detected by Northern blotting in the hypopharyngeal gland of the forager bee but not in the nurse-bee gland. These results clearly indicate that expression of the genes for these carbohydrate-metabolizing enzymes, which are needed to process nectar into honey, in the hypopharyngeal gland is associated with the age-dependent role change of the worker.

Age Factors↗

Midbrain central gray is involved in mediation of cholinergic inputs to the rostral ventrolateral medulla of the rat.

There are cholinergic inputs responsible for pressor responses in the rostral ventrolateral medulla (RVLM) and stimulation of midbrain central gray (CG) increases arterial pressure via activation of neurons in the RVLM. In this study, we examined whether the CG was involved in mediation of the cholinergic inputs to the RVLM. Male Wistar rats were anesthetized, paralyzed, and artificially ventilated. Unilateral microinjection of L-glutamate into the CG produced a pressor response. Microinjection of the muscarinic receptor antagonist scopolamine into the unilateral RVLM inhibited the pressor response to L-glutamate injected ipsilaterally into the CG, whereas microinjection of the cholinesterase inhibitor physostigmine into the RVLM enhanced it. CG stimulation also enhanced the firing rate of RVLM barosensitive neurons and the enhancement of the firing rate was inhibited by scopolamine iontophoretically applied on neurons. CG injection of L-glutamate produced a release of acetylcholine in the RVLM. Unilateral microinjection of L-glutamate into the pedunculopontine tegmental nucleus (PPT) also produced a pressor response, but the pressor response to L-glutamate was not affected by scopolamine injected ipsilaterally into the RVLM. These results provide evidence that the CG but not the PPT is involved in mediation of cholinergic inputs responsible for pressor responses in the RVLM.

Acetylcholine↗

Three novel connexin26 gene mutations in autosomal recessive non-syndromic deafness.

Since the first identification of an association between mutations in the connexin26 (Cx26) gene and autosomal recessive non-syndromic deafness it has been shown that several mutations in this gene cause recessive, sporadic, and dominant non-syndromic deafness. Three novel mutations in the Cx26 gene were identified in four of 20 Japanese families with autosomal recessive non-syndromic deafness. Seven of 40 chromosomes contained a 233delC allele, while Tyr136Stop (408C-->A) and Gly45Glu (134G-->A) were detected in two of 40 chromosomes, respectively. These mutations were not found in chromosomes in cases of sporadic congenital deafness (0/60) or in control groups (0/100). This indicates that 27.5% (11/40 chromosomes) of cases of autosomal recessive non-syndromic deafness among the Japanese are caused by mutations in the Cx26 gene.

Alleles↗

A pitch glide activates an intermediate response between auditory N1 and mismatch negativity.

Pitch glides of a continuous tone elicit auditory N1-like responses. However, their characteristics have not well been investigated, and it remained unclear whether the response is an auditory true N1 or the mismatch negativity (MMN). We found here that a rapid pitch glide activates almost the same response as a true N1. On the contrary, as the rate of the pitch glide decreases, the response continuously varies the characteristics from true N1 to MMN. This suggests that there would exist intermediate responses between auditory N1 and MMN.

Acoustic Stimulation↗

Histiocyte reaction in rabbit femurs to UHMWPE, metal, and ceramic particles in different sizes.

To assess the histologic reaction caused by biomaterial particles in different sizes around the bone-implant interface, we examined ultra-high molecular weight polyethylene (UHMWPE, average diameter of 11 microm), UHMWPE (99 microm), cobalt-chromium alloy (Co-Cr, 3.9 microm), stainless steel (SUS316L, 3.9 microm), alumina ceramics (3.9 microm), titanium alloy (Ti, 3.5 microm), Co-Cr (0.03 microm), and Ti (0.03 microm). After the longitudinal groove on a polymethylmethacrylate plug was filled with one type of the particles, the plug was inserted into the medullar canal of the distal end of rabbit femurs, and tissue block was resected 4 and 12 weeks after the insertion. Histiocytes were markedly accumulated around the particles of UHMWPE (11 microm), Co-Cr (3.9 microm), SUS316L (3.9 microm), Co-Cr (0.03 microm), and titanium alloy (0.03 microm). Around the UHMWPE particles (99 microm), a slight histiocytic reaction and bone formation were observed. Particles of alumina ceramics (3.9 microm) and titanium alloy (3.5 microm) which were in phagocytosable sizes also had few histiocytic reactions. Statistically, the material difference was more strongly related to the histiocyte reaction than to the particle size and calculated total surface area of particles. Our findings demonstrate that particles of different biomaterials and in different sizes induce different foreign-body histological reactions.

Aluminum Oxide↗

Expression of galectin-3 in fine-needle aspirates as a diagnostic marker differentiating benign from malignant thyroid neoplasms.

BACKGROUND: Galectin-3 is a beta-galactoside-binding protein that has been reported to be expressed preferentially in thyroid malignancies. The current study was designed to substantiate this finding further and to establish a presurgical diagnostic modality of differentiating between benign and malignant thyroid neoplasms by analyzing galectin-3 expression in fine-needle aspirates. METHODS: The expression of galectin-3 was examined immunohistochemically in total of 172 specimens: 45 primary and 20 metastatic papillary carcinomas, 8 primary and 2 metastatic follicular carcinomas, 5 primary and 3 metastatic anaplastic carcinomas, 3 primary medullary carcinomas, 25 follicular adenomas, 3 goiters, and 58 adjacent normal thyroid tissue. Alternatively, epithelial cells were isolated from the fine- needle aspirates of 14 thyroid nodules and subjected to immunoblotting analysis of galectin-3. RESULTS: Immunohistochemical analysis revealed that all thyroid malignancies of follicular cell origin (including papillary, follicular, and anaplastic carcinomas) showed high and diffuse expression of galectin-3, whereas one of the three medullary carcinomas of parafollicular cell origin displayed weaker and focal expression of galectin-3. In contrast, neither benign thyroid adenomas, goiters, nor normal thyroid tissues expressed galectin-3. Immunoblot analysis of the isolated epithelial cells detected galectin-3 in nine thyroid nodules that were proven histologically to be malignant ( eight papillary carcinomas and one follicular carcinoma) after surgical intervention, whereas galectin-3 was not detected in five nodules proven to be benign follicular adenomas. CONCLUSIONS: Galectin-3 serves as a marker of thyroid malignancy of follicular cell origin. Analysis of galectin-3 expression in fine-needle aspirates enhances the differential diagnostic accuracy between benign and malignant thyroid neoplasms.

Antigens, Differentiation↗

Vascular mitogen-activated protein kinase activity is enhanced via angiotensin system in spontaneously hypertensive rats.

The vascular structural remodeling function may be altered in genetically hypertensive animals, spontaneously hypertensive rats (SHR). To examine this possibility, we measured the activity of mitogen-activated protein (MAP) kinases, enzymes believed to be involved in the pathway for cell proliferation, in rat aorta strips, and examined whether the endothelium removal-induced MAP kinase activation function is altered in SHR and whether vascular angiotensin and endothelin systems are responsible for the alteration of MAP kinase activation in SHR. Male 4-week-old SHR and age-matched Wistar Kyoto rats (WKY) supplied by Charles River Japan were used. Endothelium-denuded aorta strips were incubated at 37 degrees C in medium. MAP kinase activity after incubation was time-dependently increased in strips from SHR and WKY. MAP kinase activation was greater in SHR than in WKY aorta strips. Similarly, MAP kinase activation was enhanced in aorta strips from 4-week-old SHR and stroke prone SHR supplied by the Diseases Model Cooperative Research Association (Kyoto, Japan). In aorta strips from SHR and WKY, the angiotensin receptor antagonist, losartan, and the endothelin receptor antagonist, cyclo (D-alpha-aspartyl-L-prolyl-D-valyl-L-leucyl-D-tryptophyl)(BQ123), caused concentration-dependent inhibition of MAP kinase activation. The losartan-induced but not BQ123-induced inhibition of MAP kinase activation was greater in SHR than in WKY aorta strips. Angiotensin II caused a concentration-dependent increase in MAP kinase activity and the angiotensin II-induced MAP kinase activation was greater in SHR than in WKY aorta strips. These results indicate that endothelium removal-induced MAP kinase activation is enhanced in aorta strips from young SHR, suggesting that vascular structural remodeling function may be enhanced in SHR. It appears that the enhancement of MAP kinase activation results, at least in part, from enhanced function of vascular angiotensin system in SHR.

Angiotensin II↗

A critical role for a peritumoral stromal reaction in the induction of T-cell migration responsible for interleukin-12-induced tumor regression.

Interleukin (IL) 12 has been shown to elicit tumor regression when this cytokine induces the migration of T cells to tumor sites. The present study investigates the role of a peritumoral stromal reaction in IL-12-induced T-cell migration. In the CSA1M and OV-HM tumor models, IL-12 treatment induced tumor regression that is associated with T-cell migration. Neither T-cell migration nor tumor regression was observed in the Meth A and MCH-1-A1 models. Stromal tissue containing neovascular blood vessels developed at the peritumoral area of the former two IL-12-responsive tumors but not at the peritumoral area of the latter two IL-12-unresponsive tumors. The significance of stroma development was investigated using a pair of tumor models (CSA1M and a subline derived from CSA1M designated the CSA1M variant), both of which exhibit the same tumor immunogenicity. In contrast to the parental CSA1M cell line, the variant cell line was not responsive to IL-12, and neither stroma development nor T-cell migration was observed, even after IL-12 treatment. Histological analyses revealed that the parental cell line had peritumoral stroma with intrastromal vessels but only a few vessels in tumor parenchyma, whereas the variant cell line showed no stroma but had abundant vasculature in the tumor parenchyma. Most importantly, only stromal vessels in the parental tumors expressed detectable and enhanced levels of vascular cell adhesion molecule 1 (VCAM-1)/ intercellular adhesion molecule 1 (ICAM-1) before and after IL-12 treatment, respectively. In contrast, parenchymal vasculature in the variant cell line failed to express VCAM-1/ICAM-1 even after IL-12 treatment. When transferred into recipient tumor-bearing mice, IL-12-stimulated T cells from the parental CSA1M-bearing or the variant CSA1M-bearing mice migrated into the parental but not into the variant tumor mass. Together with our previous finding that T-cell migration depends on the VCAM-1/ICAM-1 adhesive interactions, the present results indicate a critical role for peritumoral stroma/stromal vasculature in the acceptance of tumor-infiltrating T cells that is a prerequisite for IL-12-induced tumor regression.

Animals↗

Expression of mouse semaphorin H mRNA in the inner ear of mouse fetuses.

Semaphorins constitute a large family of secreted and cell-surfaced proteins that appear to function as chemorepellents to guide axons. We examined the expression pattern of M-semaH mRNA in the inner ear of mouse fetuses by in situ hybridization histochemistry. M-semaH mRNA expression was high in the endolymphatic sac involved in endolymph homeostasis. It was also high in the semicircular ducts except for the crista ampullaris, whereas no expression was detected in the epithelium of cochlear ducts.

Animals↗

Local renin-angiotensin system and mitogen-activated protein kinase activation in rat aorta.

We previously reported that endogenous angiotensin II is released to cause mitogen-activated protein (MAP) kinase stimulation in the media portion of the vasculature. In this study, we examined whether a functional renin-angiotensin system is indeed present within the media of the vasculature. In rat aortic strips, endothelium removal produced an increase of MAP kinase activity. The MAP kinase activation was inhibited either by the renin inhibitor pepstatin A or by the angiotensin-converting enzyme inhibitor captopril. The degree of the inhibition of the MAP kinase activation by pepstatin A, captopril and the angiotensin receptor antagonist losartan was almost the same. Pepstatin A inhibited MAP kinase activation induced by renin but not by angiotensin I and angiotensin II. Captopril inhibited the MAP kinase activation induced by angiotensin I but not by angiotensin II. In nephrectomized rat aortic strips, endothelium removal also produced an increase in MAP kinase activity, but the MAP kinase activation was considerably small and minimally inhibited by losartan. Nephrectomy produced a marked decrease in plasma renin activity. These findings suggest that an apparently fully intact and functional renin-angiotensin system is present in the media of the rat vasculature and this system serves to increase MAP kinase activity. It appears that renin plays the determining role in the regulation of angiotensin generation also in the media and the major source of the renin is renin of kidney origin.

Angiotensin I↗

Nitric oxide in the flocculus works the inhibitory neural circuits after unilateral labyrinthectomy.

We previously reported that nitric oxide (NO) production in the unipolar brush (UB) cells is involved in vestibular compensation [T. Kitahara, N. Takeda, P.C. Emson, T. Kubo, H. Kiyama, Changes in nitric oxide synthase-like immunoreactivities in unipolar brush cells in the rat cerebellar flocculus after unilateral labyrinthectomy, Brain Res. 765 (1997) 1-6]. To further elucidate the role of NO-mediated signaling in flocculus after unilateral labyrinthectomy (UL), we examined UL-induced Fos expression, a marker of neural activity, in vestibular brainstem with continuous floccular infusions of Nomega-nitro-l-arginine methyl ester (l-NAME), an inhibitor of NO synthase (NOS). After UL with floccular l-NAME infusions, Fos expression appeared in bilateral medial vestibular (MVe) and prepositus hypoglossal (PrH) nuclei. After UL with floccular saline infusions, however, Fos expression was observed only in the ipsi-MVe and contra-PrH. Furthermore, it has been revealed that UL with l-NAME infusions caused more severe vestibulo-ocular disturbances than UL with saline infusions at the initial stage [Kitahara et al. Brain Res. 765 (1997) 1-6]. Therefore, it is suggested that UL with floccular l-NAME infusions activates the contra-MVe and ipsi-PrH neurons and causes more severe imbalance between intervestibular nuclear activities at the initial stage. NO-mediated signaling in flocculus could be a possible driving force of the flocculus-mediated inhibition on the contra-MVe and ipsi-PrH at the initial stage of vestibular compensation.

Animals↗

Prenatal diagnosis of long QT syndrome using fetal magnetocardiography.

We describe the detection of congenital long QT syndrome in a fetus at 37 weeks' gestation using magnetocardiography (MCG). The prenatal diagnosis was confirmed by standard electrocardiography (ECG) performed after birth. This is the first case report of fetal long QT syndrome detected by MCG. Fetal MCG may be useful in the prenatal diagnosis of congenital cardiac disease with abnormal ECG findings.

Adult↗

Expression of major urinary protein genes in the nasal glands associated with general olfaction.

Gene expression of major urinary protein (MUP) mRNAs was examined in the mouse nasal tissue. By polymerase chain reaction, we identified two cDNA segments encoding MUP 4 and MUP 5 genes in the nose. The expression level of both MUP 4 and 5 mRNAs in the nasal tissue was very high and exceeded that of the liver. Liver MUPs are excreted into the urine and are known to play an important role in pheromonal communication. We showed that nose and liver MUPs were composed of different subtypes of MUPs and that nose MUP mRNAs was detected in prepubescent periods when liver MUP mRNAs had not yet been transcripted. In situ hybridization revealed that nose MUP mRNAs are localized in the lateral wall and nasal septum and their expression pattern is identical to that of rat odorant-binding protein (OBP)-I. We also identified cDNA of mouse OBP-II gene from the nasal tissue and showed that the expression pattern of MUP gene was identical to that of OBP-II gene in the nose. These histological data indicate that nose MUPs are favorable for catching odorant molecules rather than pheromones, and may share their function with OBPs.

Amino Acid Sequence↗

A clinicopathologic study of transient osteoporosis of the hip.

OBJECTIVE: It has been proposed that transient osteoporosis of the hip (TOH) may represent the early reversible phase of osteonecrosis of the femoral head (ON). The purpose of this study was to investigate the clinicopathologic characteristics of three cases of TOH. DESIGN AND PATIENTS: A bone biopsy was performed on three patients who had been diagnosed as having TOH based on the clinical course, radiograph, bone scintigram, and MR images. The biopsy specimens were studied histopathologically by light and electron microscopy. RESULTS: The most characteristic feature of TOH was focal areas of thin and disconnected bone trabeculae covered by osteoid seams and active osteoblasts. The surrounding bone marrow tissue showed edematous changes and mild fibrosis, frequently associated with vascular congestion and/or interstitial hemorrhage. No osteonecrotic region was observed in either the bone trabeculae or the bone marrow tissue. All patients have improved clinically and in the 3.5-9 years of follow-up have shown no evidence of ON. CONCLUSIONS: This study supports the concept that transient osteoporosis of the hip is a distinct entity.

Adult↗