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Biomedical subjects

K Kikuchi

Publications and source records attributed to K Kikuchi.

At least 19 recordsLinked to original sources

Cytoplasmic glutathione regulated by cumulus cells during porcine oocyte maturation affects fertilization and embryonic development in vitro.

It is generally accepted that cumulus cells support the nuclear maturation of mammalian oocytes. In the present study, we examined relationships between the cytoplasmic glutathione (GSH) content of porcine oocytes, and oocyte nuclear maturation, fertilization or subsequent embryonic development. Cumulus-oocyte complexes (COCs; control group) and oocytes denuded of cumulus cells after collection (DO 0h group) were cultured for 24h with dibutyryl cAMP, eCG and hCG (first culture step) and then for a further 20h without supplements (second culture step; 44h total culture). After the first culture step, some of the COCs were denuded, either completely (DO 24h group) or partly (H-DO 24h group), and then matured by the second culture step. Also, in the second culture step, some DOs were co-cultured with cumulus cells that had been pre-cultured for 24h (DO 24h+CC group). The maturation rates of all the cumulus-removed groups (DO 0h, DO 24h, H-DO 24h and DO 24h+CC groups) were lower (34.3-45.0%) than that of the control group (64.5%; P<0.05). The GSH contents of matured oocytes in the completely denuded groups (DO 0h, DO 24h and DO 24h+CC groups) were lower (4.03-5.26pmol/oocyte) than that of the control group (9.60pmol/oocyte; P<0.05); however, the H-DO 24h group had an intermediate value (7.0pmol/oocyte). The male pronuclear formation rates of completely denuded oocytes were lower (41.4-59.3%) than that of the control group (89.4%; P<0.05), whereas the H-DO 24h group had an intermediate rate (80.0%). The blastocyst formation rates of the completely denuded oocytes were lower (3.0-4.5%) than that of the control group (19.9%; P<0.05), and the H-DO 24h group again had an intermediate rate (11.6%). The GSH content was correlated with the rates of male pronuclear formation (P<0.01) and blastocyst formation (P<0.01), and also with the number of cells per blastocyst (P<0.01). In conclusion, we inferred that GSH synthesized by intact cumulus cells during maturation culture improved oocyte maturation and played an important role in fertilization and embryonic development.

Animals↗

Secretory IgA, salivary peroxidase, and catalase-mediated microbicidal activity during hydrogen peroxide catabolism in viridans streptococci: pathogen coaggregation.

Viridans streptococci can kill methicillin-resistant Staphylococcus aureus (MRSA) through the production of hydrogen peroxide (H2O2). However, several hundred viridans streptococci cells are necessary to kill 1 cfu of MRSA. We analyzed the potency of bactericidal and fungicidal effector molecules induced by catabolism of H2O2 in the oral cavity. Secretory IgA (SIgA) and an unidentified salivary component bound Streptococcus sanguinis, a viridans streprococcus, and MRSA into coaggregates. In these coaggregates, salivary peroxidase and the MRSA catalase produced singlet molecular oxygen (1O2) from H2O2 produced by viridans streptococci. SIgA converted 1O2 into ozone, which has potent bactericidal and fungicidal activity. We calculated that <10 cfu of Streptococcus sanguinis were necessary to kill 1 cfu of MRSA in the coaggregate. SIgA, Aspergillus niger catalase, and H2O2 in saliva killed Candida albicans, which is highly resistant to reagent H2O2. Together with indigenous bacteria and innate immunity, SIgA potentially constitutes a novel system that may sustain oral homeostasis.

Adult↗

Pseudoscleroderma associated with cancer.

In the period 1994-2002, we saw 71 patients with sclerotic skin changes; 66 were diagnosed with systemic sclerosis (SSc) while five (7%) were diagnosed with pseudoscleroderma associated with various malignancies. The mean duration of disease in these five patients was significantly shorter than that of patients with SSc. The incidence of positive antinuclear antibodies, Raynaud's phenomenon or oesophageal involvement in patients with pseudoscleroderma was significantly lower than that in patients with SSc. The distributions of skin sclerosis varied in each case. Serum basic fibroblast growth factor (bFGF) levels in the five patients with pseudoscleroderma were very elevated compared with levels in controls. Elevated expression of bFGF was detected on fibroblasts of affected skin and in one lung-cancer tissue sample obtained by excision.

Adenocarcinoma↗

Coexpression of an unusual form of the EWS-WT1 fusion transcript and interleukin 2/15 receptor betamRNA in a desmoplastic small round cell tumour.

BACKGROUND: The beta chain of the interleukin 2/15 receptor (IL-2/15Rbeta) is induced by the expression of the EWS-WT1. A case of desmoplastic small round cell tumour (DSRCT) expressing only an unusual EWS-WT1 treated by us is reported here. AIM: To characterise an unusual form of EWS-WT1. METHODS: Frozen tissue sections of the axillary tumour were examined using a laser-assisted microdissection technique and reverse transcriptase polymerase chain reaction. RESULTS: The novel fusion of exon 8 of EWS and the defective exon 10 of WT1 (-KTS) was detected. Although it was an unusual form, the coexpression of the present EWS-WT1, IL-2/15Rbeta and Janus kinase (JAK1) mRNA was detected in the tumour cells. IL-2 and signal transducers and activators of transcription (STAT5) mRNA were detected in both tumour and stromal cells. CONCLUSION: The induction of the IL-2/15 receptor signalling pathway may contribute to tumorigenesis in DSRCT through a paracrine or an autocrine system, even though the EWS-WT1 was an unusual form.

Adult↗

[Surgical resection for sternal metastasis from gastric cancer; report of a case].

A 72-year-old woman underwent a distal gastrectomy for gastric cancer 8 years ago. Histopathological examination showed poor differentiated adenocarcinoma. She had no symptom but chest computed tomography (CT) revealed tumor at sternum. A percutaneous needle biopsy under CT was performed and showed poor differentiated adenocarcinoma. Therefore, extirpation was performed. The tumor was located in sternum and left 2nd rib and thought to invade greater pectoral muscle and left lung. The tumor was resected at sufficient margin with surrounding tissues. Compared with tissue of gastric cancer for histopathological examination, sternal tumor had homeomorphous formation and was diagnosed sternal metastasis from gastric cancer. Sternal metastasis from gastric cancer is a rare disease and standard therapy is controversial. In this case, no findings of reccurrence have been found for 8 months after the operation.

Adenocarcinoma↗

[Aortic valve replacement for calcified aortic valve and annulus with ultrasonic decalcification].

UNLABELLED: We conducted ultrasonic decalcification on calcified annulus in patients with aortic stenosis (AS) using an ultrasonic operator, Sonopet (UST 2001) prior to aortic valve replacement (AVR). We studied the reliability of this method. SUBJECT AND METHOD: From January 2002 to August 2005, AVR was conducted for AS using the Sonopet in 45 patients, comprising of 18 male and 27 female subjects. The mean age was 73.3 +/- 9.7. RESULT: Artificial valves were successfully inserted at the intra-annular level in 37 patients and at the supra-annular level in 8 patients without conducting annular enlargement. In the patients with narrow annuli of less than 19 mm (23 patients), the preoperative mean annular diameter was 18.2 +/- 1.0 mm, but significantly larger artificial valves with an average diameter of 19.3 +/- 1.5 mm (p=0.003) were successfully inserted. CONCLUSION: AVR was proved to be safe and easy by previous ultrasonic decalcification of the annuls using the Sonopet. This method was very useful because it required no enlargement of aortic annulus.

Aged↗

[Off-pump coronary artery bypass in high-risk patients].

In this study, 110 consecutive patients who had undergone off-pump coronary artery bypass (OPCAB) in the past 2 years were evaluated for early results of OPCAB. Patients were classified as a high-risk group (H group: 68 patients consisting of 46 men and 22 women) and a low-risk group (L group: 42 patients consisting of 31 men and 11 women), respectively, and were evaluated for the early operative results. No differences were noted between the H and L groups in the mean number of bypass grafts (2.9 +/- 0.9 in the H group, 2.9 +/- 0.9 in the L group), the rates of complete revascularization (85% in the H group, 93% in the L group), those of various graft materials bypassed, or those of sequential bypass. In all patients, we were able to undergo coronary revascularization by the aortic no-touch technique using arterial grafts exclusively. In the H group, 1 patient (1.5%) died in hospital, but no patients developed cerebral infarction postoperatively, and the frequency of complications was similar to that in the L group. The results of OPCAB for high-risk patients were good, and it was suggested that OPCAB using in situ arterial grafts was very useful particularly in patients with cerebrovascular diseases.

Aged↗

Participation of the nonreducing terminal beta-galactosyl residues of the neutral N-linked carbohydrate chains of porcine zona pellucida glycoproteins in sperm-egg binding.

The zona pellucida (ZP) surrounding the mammalian oocyte is composed of three glycoprotein components (ZPA, ZPB, and ZPC). Mammalian sperm bind to carbohydrate chains of a ZP glycoprotein in the initial phase of fertilization. Sperm-ligand carbohydrate chains have been characterized in mouse, cow, and pig. In pigs, triantennary/tetraantennary neutral complex-type chains from ZPB/ZPC mixture possess stronger sperm-binding activity than those of biantennary chains (Kudo et al., 1998: Eur J Biochem 252:492-499). Most of these oligosaccharides have beta-galactosyl residues at the nonreducing ends. This study used two in vitro competition assays to investigate the participation of the nonreducing terminal beta-galactosyl residues of the ligand active chains in porcine sperm binding. The removal of the nonreducing terminal beta-galactosyl residues from either the ligand active carbohydrate chains or endo-beta-galactosidase-digested glycoproteins significantly reduced their inhibition of sperm-egg binding, indicating that the beta-galactosyl residues at the nonreducing ends are involved in porcine sperm-egg binding. A correlation between the sperm-binding activity and in vitro fertilization rate is also presented.

Animals↗

Large-scale DNA microarray analysis of atopic skin lesions shows overexpression of an epidermal differentiation gene cluster in the alternative pathway and lack of protective gene expression in the cornified envelope.

BACKGROUND: Atopic dermatitis (AD)-specific genes have not yet been clarified. Objectives To identify gene expression specific to active atopic skin lesions. METHODS: We analysed 23,000 genes in skin biopsy samples from 17 patients with AD and four normal controls using Affymetrix oligonucleotide arrays. RESULTS: Four of the 10 genes with the greatest differences in expression between patients and controls, S100A8 and S100A7 (upregulated), and loricrin and filaggrin (downregulated), were epidermal differentiation genes located on 1q21, a locus previously reported to have a genetic linkage with AD. CONCLUSIONS: Our results, showing downregulation of the cornified envelope genes and upregulation of the alternative keratinization pathway, are the first to suggest abnormal epidermal differentiation and defective defences as key abnormalities in AD.

Adult↗

Functional properties of the stratum corneum in patients with diabetes mellitus: similarities to senile xerosis.

BACKGROUND: Although diabetes mellitus is known to induce many pathophysiological changes in the skin, none of the earlier studies has focused on the state of the stratum corneum (SC) in patients with diabetes. In our previous report on a diabetic mouse model, we described a decreased SC hydration state and abnormalities in the function of the sebaceous gland together with a normally retained barrier function of the SC. OBJECTIVES: To examine the functional changes of the SC in patients with diabetes. METHODS: The following values were measured in 49 patients with diabetes: fasting plasma glucose (FPG) and haemoglobin A1c (HbA1c); skin surface lipid level on the forehead; and high-frequency conductance (HFC) and transepidermal water loss (TEWL) of the SC of the volar forearm and extensor surface of the lower leg. These functions of the SC were compared in two pairs of patient groups, i.e. between groups with high and low FPG, and between groups with high and low HbA1c. RESULTS: The HFC levels on the extensor leg and volar forearm were significantly lower in the group with high FPG (FPG > 110 mg dL(-1)) than in the group with low FPG (FPG < 110 mg dL(-1)), although TEWL values were almost the same in these two groups. The skin surface lipids on the forehead were significantly lower in the former than in the latter. The HFC did not differ significantly between the group with high HbA1c (HbA1c > 5.8%) and the group with low HbA1c (HbA1c < 5.8%), whereas the TEWL of the volar forearm was slightly lower in the former than in the latter. CONCLUSIONS: These new results as well as our earlier results with diabetic mice suggest that patients with diabetes mellitus tend to show a reduced hydration state of the SC together with decreased sebaceous gland activity, without any impairment of the SC barrier function.

Adolescent↗

Doxapram inhibits carotid sinus baroreceptors in rabbits.

1 To clarify the effects of doxapram on the baroreflex, we recorded carotid sinus nerve (CSN) activity in isolated and perfused carotid artery bifurcations of rabbits. 2 The CSN activity due to chemoreceptor stimulation was blocked by resection of the nerve branches from the carotid body. After the resection, the CSN activity was correlated to increase of carotid sinus (CS) pressure. 3 Administration of doxapram reduced the CSN activity originating from baroreceptors. The effect of doxapram on baroreceptors was dose dependent and reversible. 4 It is unlikely that doxapram altered CS wall mechanics because CS pressure did not change in the presence of the drug. 5 We conclude that doxapram acts on the cardiovascular system in part by inhibiting the negative feedback loop that originates in CS baroreceptors.

Animals↗

The expression of platelet-derived growth factor and connective tissue growth factor in different types of abdominal aortic aneurysms.

AIM: An atherosclerotic abdominal aortic aneurysms (AAAA) differ from inflammatory abdominal aortic aneurysms (IAAA), which are characterized by a non specific inflammatory reaction leading to considerable aneurysmal wall thickness from the media to adventitia and retroperitoneal fibrosis in the surrounding tissue. Platelet-derived growth factor (PDGF) and its receptor have been localized to specific cell types within atherosclerotic plaques. Human connective tissue growth factor (CTGF) is a cysteine rich polypeptide that has similar structures to PDGF and has been implicated in connective tissue formation. PDGF and CTGF may play a role in the development of aneurysmal walls in both AAAA and IAAA. METHODS: Using in situ hybridization technique with DIG-labeled RNA probes and immunostaining, we investigated CTGF gene expression, and expression of PDGF and its receptor protein, in human aneurysmal walls. RESULTS: Expression of CTGF mRNA was found on vascular smooth muscle cells (VSMC) in specimens from AAAA and IAAA. Strong CTGF expression was localized in VSMC around calcification in AAAA. In IAAA, strong expression of CTGF was observed around inflammatory cells. In the aneurysmal walls of AAAA, PDGF A and B chains were strongly stained on small vessels, and the PDGF beta receptor was also strongly stained on VSMC around calcification. In the aneurysmal wall of IAAA, weak expressions of PDGF A and B chains were observed in endothelial cells of vessel walls around the inflammatory cells, but the intensity of expression was much weaker than that on the vessel walls in AAAA. CONCLUSIONS: Such differences in fibrogenic cytokine expression may be involved in characteristic aneurysmal formation.

Adult↗

The dural tail of intracranial meningiomas on fluid-attenuated inversion-recovery images.

We evaluated the "dural tail" associated with 48 intracranial meningiomas on fluid-attenuated inversion-recovery (FLAIR) and contrast-enhanced T1-weighted images. In 30 (62.5%), a dural tail was observed on contrast-enhanced T1-weighted images, and thickening of the dura mater and abnormal signal were identified in the corresponding region on FLAIR images. Thus, FLAIR imaging was useful for showing dural abnormality associated with meningiomas without the needed for contrast medium.

Adult↗

Distinct glutamate receptors govern differential levels of nitric oxide production in a layer-specific manner in the rat cerebellar cortex.

To evaluate roles of nitric oxide (NO) in neural functions, it is critical to know how neural inputs activate neuronal NO synthase in individual sites. Although NMDA receptor-dependent mechanism well explains postsynaptic, robust NO production, this sole mechanism does not explain some aspects of NO production in the brain, such as the low-level production of NO and the mechanism for presynaptic NO production. We hypothesized that the glutamate receptor involved in NO production is site-specific and controls the initial NO concentration in each site. We visualized NO production mediated by NMDA, AMPA and type-1 metabotropic glutamate (mGlu-1) receptors in rat cerebellar slices and granule cells in culture, with an NO-specific fluorescent indicator, diaminofluorescein-2. AMPA receptor, but not NMDA or mGlu-1 receptor, was responsible for NO production at parallel fiber terminals, which was blocked by CNQX, tetrodotoxin or voltage-dependent calcium channel blockers. More numbers of electrical stimulation were required for NO production in the molecular layer than in other layers, suggesting that AMPA receptor activation generates NO at lower concentrations through a remote interaction with NO synthase. Although Purkinje cell does not express NO synthase, we detected NO production in Purkinje cell layer following electrical stimulation in the white matter at 50 Hz, but not at 10 Hz. This NO production was tetrodotoxin-sensitive, suggesting occurrence in the basket cell terminals, and required synergistic activation of mGlu-1 and NMDA receptors. In the granule cell layer, activation of AMPA or mGlu-1 receptor produced NO uniformly, while NMDA receptor activation produced NO in discontinuous areas of this layer. Thus, distinct glutamate receptors, including non-NMDA receptors, govern occurrence and level of NO production in a layer-specific manner.

Anesthetics, Local↗