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Biomedical subjects

Katsuyuki Kinoshita

Publications and source records attributed to Katsuyuki Kinoshita.

At least 19 recordsLinked to original sources

IL-4Ralpha polymorphism in regulation of IL-4 synthesis by T cells: implication in susceptibility to a subset of murine lupus.

To thoroughly understand the role of IL-4 in the pathogenesis of systemic lupus erythematosus (SLE), a prototypic antibody-mediated systemic autoimmune disease, we examined the potential of in vitro IL-4 production by anti-CD3 mAb-stimulated splenic T cells in SLE model of NZB, BXSB and related mouse strains. Unexpectedly, both SLE-prone NZB and BXSB mice had a limited potential to produce IL-4, while disease-free NZW mice had a high potential. Levels in (NZB x NZW) F1 and (NZW x BXSB) F1 were in between. Genome-wide search for quantitative trait loci (QTL) controlling this variation identified a single significant QTL in the vicinity of IL-4Ralpha gene on chromosome 7. Sequence analysis of IL-4Ralpha cDNA revealed that there are 17 nucleotide substitutions resulting in eight amino acid changes between NZB and NZW strains. BXSB showed the identical sequence, as did NZB. Thus, it was suggested that the NZW-type polymorphism controls a high potential and the NZB/BXSB-type polymorphism controls a low potential for IL-4 production by T cells. Linkage studies using NZW x (NZW x BXSB) F1 male and (NZB x NZW) F1 x NZW female back-cross mice revealed that the BXSB/NZB-type IL-4Ralpha polymorphism significantly linked to BXSB, but not to (NZB x NZW) F1 lupus. Thus, the low IL-4-producing phenotype appears to predispose to SLE in BXSB, but not NZB-related strains, suggesting that the role of IL-4 in the pathogenesis may differ between certain subsets of SLE, even if they show similar disease phenotypes.

Animals↗

Genetic dissection of the effects of stimulatory and inhibitory IgG Fc receptors on murine lupus.

Immune complex (IC)-mediated tissue inflammation is controlled by stimulatory and inhibitory IgG Fc receptors (FcgammaRs). Systemic lupus erythematosus is a prototype of IC-mediated autoimmune disease; thus, imbalance of these two types of FcgammaRs is probably involved in pathogenesis. However, how and to what extent each FcgammaR contributes to the disease remains unclear. In lupus-prone BXSB mice, while stimulatory FcgammaRs are intact, inhibitory FcgammaRIIB expression is impaired because of promoter region polymorphism. To dissect roles of stimulatory and inhibitory FcgammaRs, we established two gene-manipulated BXSB strains: one deficient in stimulatory FcgammaRs (BXSB.gamma(-/-)) and the other carrying wild-type Fcgr2b (BXSB.IIB(B6/B6)). The disease features were markedly suppressed in both mutant strains. Despite intact renal function, however, BXSB.gamma(-/-) had IC deposition in glomeruli associated with high-serum IgG anti-DNA Ab levels, in contrast to BXSB.IIB(B6/B6), which showed intact renal pathology and anti-DNA levels. Lymphocytes in BXSB.gamma(-/-) were activated, as in wild-type BXSB, but not in BXSB.IIB(B6/B6). Our results strongly suggest that both types of FcgammaRs in BXSB mice are differently involved in the process of disease progression, in which, while stimulatory FcgammaRs play roles in effecter phase of IC-mediated tissue inflammation, the BXSB-type impaired FcgammaRIIB promotes spontaneous activation of self-reactive lymphocytes and associated production of large amounts of autoantibodies and ICs.

Animals↗

The role of X/Y linker region and N-terminal EF-hand domain in nuclear translocation and Ca2+ oscillation-inducing activities of phospholipase Czeta, a mammalian egg-activating factor.

Sperm-specific phospholipase C-zeta (PLCzeta) causes intracellular Ca(2+) oscillations and thereby egg activation and is accumulated into the formed pronucleus (PN) when expressed in mouse eggs by injection of cRNA encoding PLCzeta, which consists of four EF-hand domains (EF1-EF4) in the N terminus, X and Y catalytic domains, and C-terminal C2 domain. Those activities were analyzed by expressing PLCzeta mutants tagged with fluorescent protein Venus by injection of cRNA into unfertilized eggs or 1-cell embryos after fertilization. Nuclear localization signal (NLS) existed at 374-381 in the X/Y linker region. Nuclear translocation was lost by replacement of Arg(376), Lys(377), Arg(378), Lys(379), or Lys(381) with glutamate, whereas Ca(2+) oscillations were conserved. Nuclear targeting was also absent for point mutation of Lys(299) and/or Lys(301) in the C terminus of X domain, or Trp(13), Phe(14), or Val(18) in the N terminus of EF1. Ca(2+) oscillation-inducing activity was lost by the former mutation and was remarkably inhibited by the latter. A short sequence 374-383 fused with Venus showed active translocation into the nucleus of COS-7 cells, but 296-309 or 1-19 did not. Despite the presence of these special regions, both activities were deprived by deletion of not only EF1 but also EF2-4 or C2 domain. Thus, PLCzeta is driven into the nucleus primarily by the aid of NLS and putative regulatory sites, but coordinated three-dimensional structure, possibly formed by a folding in the X/Y linker and close EF/C2 contact as in PLCdelta1, seems to be required not only for enzymatic activity but also for nuclear translocation ability.

Active Transport, Cell Nucleus↗

Does estrogen affect the development of abnormal vascular function in offspring of rats fed a low-protein diet in pregnancy?

It is established that there are gender-related differences in the effects on offspring blood pressure induced by maternal protein restriction in animal studies. Since such effects may depend on estrogen levels, we hypothesized that lower estrogen would induce an earlier onset of hypertension caused by maternal under-nutrition. Wistar rats were fed a diet containing either 18% (C) or 9% (R) casein throughout pregnancy. Half of the offspring in both C and R groups were ovariectomized on day 50 (CX, RX), and the other half underwent a sham operation (CO, RO). On d 175, offspring were killed for small artery reactivity and histologic investigation. Birth weight and later growth were not significantly different between C and R. RX had higher systolic blood pressure than CX on d125, but no difference was seen between RO and CO. On d 175, systolic blood pressure was higher in R than in C, whether or not ovariectomized. Dilator responses to acetylcholine and bradykinin in small mesenteric arteries were significantly attenuated in RX, although responses to SNP and isoprenaline showed no attenuation in R. The ratio of coronary peri-vascular fibrosis to total vascular area was higher in R, and the fibrosis became prominent in ovariectomized rats. These findings suggest that estrogen plays an important role in limiting the elevation of offspring blood pressure induced by maternal under-nutrition, possibly via BK-mediated mechanisms. The processes may underlie gender and life course patterns of hypertension and also the developmental origins of this disease.

Animals↗

Hemodynamic evaluation of a prenatal thoracoamniotic shunt for fetal pleural effusion.

AIMS: To evaluate the efficacy of a thoracoamniotic shunt for the treatment of pleural effusion (PE) in the view of hemodynamics. METHODS: The preload index (PLI) in the inferior vena cava (IVC), the maximal flow velocity of the descending aorta (VAomax), skin edema on the thorax and the ratio of lung to the thorax transverse area (L/T) as measured by ultrasound were evaluated before and after thoracoamniotic shunt placement for 5 fetuses with PE. RESULTS: The PLI and skin edema on the thorax decreased significantly after shunt placement compared to before shunt placement (PLI before: 0.488 +/- 0.036, after: 0.348 +/- 0.043, P < 0.05; edema before: 15.3 +/- 2.06 mm, after: 9.00 +/- 0.63 mm, P < 0.05). Furthermore, the L/T increased significantly after shunt placement compared to before (before: 0.220 +/- 0.013, after: 0.260 +/- 0.011, P < 0.01). No significant difference in VAomax was seen between before and after shunt placement (before: 101.5 +/ -6.39 cm/s, after: 10.7.6 +/ -5.41 cm/s, P = 0.16). CONCLUSIONS: The shunt for PE improved PLI especially in the fetal hemodynamics significantly.

Adult↗

Changes in matrix metalloproteinase 2 activities in amniochorions during premature rupture of membranes.

OBJECTIVE: Increased proteolytic activities of matrix metalloproteinases (MMPs) such as MMP-3 and MMP-9 are associated with premature rupture of membranes at term. However, it is unclear whether MMP-2 is involved in the premature rupture of membranes. In this study, to elucidate the role of MMP-2, we evaluated the activity of MMP-2 and also the expression of pro-MMP-2, membrane type 1 (MT1)-MMP and tissue inhibitor of metalloproteinase (TIMP)-1 in premature rupture of membranes. METHODS: Amniochorions were prepared from 29 subjects with no labor (cesarean section; CS, n = 10), labor (normal delivery; ND, n = 10), and labor during premature rupture of membranes (PROM, n = 9). MMP-2 activity was spectrophotometrically assayed by measuring the digestion of an MMP-2-specific substrate. The levels of pro-MMP-2, MT1-MMP and TIMP-1 were determined by Western immunoblotting. RESULTS: The activity of MMP-2 in PROM was significantly higher than that in CS and ND (P <.05). In addition, the levels of MT1-MMP, an activator of MMP-2, were higher in PROM than in CS and ND. In contrast, the level of TIMP-1, an inhibitor of MMP-2 was substantially lower in PROM than CS and ND. Moreover, the levels of pro-MMP-2 were increased more significantly in PROM and ND than in CS (P <.05). CONCLUSION: Our results suggest that the increased expression of pro-MMP-2 and MT1-MMP and decreased expression of TIMP-1 may result in the increased activity of MMP-2, which is involved in the degradation of extracellular matrix (ECM) of fetal membrane, thereby inducing the premature rupture of membranes at term.

Amnion↗

Molecular characterization of a germ-cell-specific antigen, TEX101, from mouse testis.

TEX101, a glycoprotein we recently identified, is primarily characterized as a unique germ-cell-specific marker protein that shows sexually dimorphic expression during mouse gonad development. Based on data obtained from molecular biological as well as immuno-morphological studies, we believe this molecule may play a role in the process underlying germ cell formation. However, many points remain unclear as the molecular characteristics and its physiological functions are far from being completely understood. To clarify the molecular basis of TEX101, we herein report a further biochemical characterization of the molecule using testicular Triton X-100 extracts from mice. Deglycosylation studies using endoglycohydrolases that delete N-linked oligosaccharides (OS) from the molecule show that TEX101 is highly (approximately 47%) N-glycosylated. All potential N-glycosylation sites within TEX101 are glycosylated and most of these sites are occupied by endoglycosidase F2-sensitive biantennary complex type OS units. In addition, an extremely low population among TEX101 possesses only endoglycosidase H-sensitive hybrid type OS units. In studies using phosphatidylinositol-specific phospholipase C against native testicular cells or TEX101 transfectant, the enzyme treatment caused major reduction of the TEX101 expression on the cell, suggesting that TEX101, at least in part, is expressed as a glycosylphosphatidylinositol-anchored protein. Taken together, these findings will help elucidate the molecular nature of TEX101, a marker molecule that appeared on germ cells during gametogenesis.

Animals↗

A novel method of ninth-intercostal microlaparoscopic approach for patients with previous laparotomy.

BACKGROUND: To evaluate the availability of a primary laparoscopic approach for a patient with previous laparotomy and to assess postoperative adhesion to the abdominal wall.. METHODS: A total of 172 patients with a history of laparotomy who were undergoing laparoscopic surgery in our hospital were evaluated. The primary trocar insertion was performed via the ninth-intercostal microlaparoscopic approach for these patients. RESULTS: This procedure could be performed on all patients and there were no complications. Postoperative abdominal wall adhesion was found in 53 (30.8%) of 172 patients, and periumbilical adhesion was found in 16 patients (9.3%). Among the 172 patients, 150, 19, and three patients had undergone laparotomy once, twice, and thrice, respectively, prior to this study. Considering the number of previous laparotomies, the frequency of periumbilical adhesion was 5.3% (eight of 150 patients), 36.8% (seven of 19 patients), and 33.3% (one of three patients), respectively. Among 150 patients who had undergone laparotomy once, the type of laparotomy was an obstetric-and-gynecologic surgery in 126 patients and other surgeries in 24 patients. There was no significance in the frequency of adhesion between types of previous laparotomies. Surgical incisions were classified as median infra-umbilical incision (94 patients), median supra-umbilical incision (three patients), Pfannenstiel's incision (33 patients), para-rectal incision (15 patients), and peri-rectal incision (five patients). A high frequency of periumbilical adhesion was recognized in the all three median supra-umbilical incisions. CONCLUSION: Ninth-intercostal microlaparoscopic approach is safe for laparoscopic surgery in patients who have undergone laparotomy previously, and this procedure could prevent the risk of bowel injury.

Abdominal Wall↗

Recurrence rate of endometriomas following a laparoscopic cystectomy.

BACKGROUND: The objectives were to observe the clinical course of patients who underwent laparoscopic cystectomy (LC) using transvaginal ultrasonography. METHOD: The definition of "recurrence" was to detect a diffuse, hypoechoic area whose long axis was at least 2 centimeters (cm) on the transvaginal ultrasonogram during the postoperative follow-up period. Using the Cox regression test, we analyzed each patient's background variables, namely, patient's age, the revised American Society of Reproductive Medicine score (r-ASRM score), use of gonadotropin releasing hormone analogues (GnRHa), and pregnancy during the observation period. Next, cyst size and multi-lobularity of each of the 417 cysts were also analyzed as recurrence risk factors per cyst. RESULTS: The mean postoperative observation period was 21.4+/-16.8 months. During the observation period, 50 (15.9%) out of 315 patients experienced recurrence and 51 (16.2%) out of 315 patients became pregnant. The cumulative recurrence rate per patient was 31.7% over 60 months. Among prognostic factors per patient, age and showed negative and r-ASRM scores positive correlations with cyst recurrence. On the other hand, we did not identify any cyst factors associated with recurrence. The patients who underwent hemilateral LC showed a 5.2% cyst recurrence rate in the ovary that appeared to be normal at the time of the initial surgery. CONCLUSION: Young age and severe endometriosis appeared to be the factors associated with high recurrence risk. Recurring ovarian endometrimas probably include cysts occurring spontaneously and those recurring from the cyst residues in the lesions where cystectomy has been performed.

Adolescent↗

Simultaneous laparoscopic treatment for rectosigmoid and ileal endometriosis.

Endometriosis is common in women of childbearing age, while severe intestinal endometriosis requiring bowel resection is relatively rare. Intestinal endometriosis has recently been managed laparoscopically. We report the case of a 38-year-old patient with rectosigmoid and ileal endometriosis who was successfully treated by laparoscopic bowel resections. The patient had first presented at age 34 years with a chief complaint of rectal bleeding and lower abdominal pain related to the menstrual cycle. She underwent laparoscopic surgery and was diagnosed with severe endometriosis involving the rectosigmoid colon. Although an additional laparoscopic surgery had been planned, she did not return to the hospital. When she was 38 years old, she presented again with the same symptoms. Magnetic resonance imaging revealed a low intensity mass between the uterus and the rectosigmoid colon. A barium enema showed a stenotic site in the rectosigmoid colon. After hormone therapy, she underwent laparoscopic surgery. The anterior wall of the rectosigmoid colon adhered firmly to the corpus of the uterus, and another stenotic site was identified at the terminal ileum. The rectosigmoid colon and ileum were partially resected under laparoscopy. The postoperative course was uneventful and she was freed of symptoms. Laparoscopic treatment for patients with severe endometriosis of the bowel has becomes feasible and safe.

Adult↗

A novel instrument and technique for using Seprafilm hyaluronic acid/carboxymethylcellulose membrane during laparoscopic myomectomy.

Myomectomy is associated with the development of adhesions, and even when the patient is operated on laparoscopically, postoperative adhesions can often occur. Seprafilm is an effective and highly safe barrier to postoperative adhesions. However, it is vulnerable to tears and difficult to handle. We developed a device to introduce Seprafilm into the abdominal cavity during the laparoscopic procedure. Seprafilm is divided into 6 pieces and the pieces are rolled and inserted into the purpose-built introducer to be guided into the abdominal cavity. The inner syringe is pushed in and Seprafilm is applied onto the uterine wound with two grasping forceps. Of 117 patients in our study, we were able to use Seprafilm on 114 patients (97.4%). In total, 221 pieces of Seprafilm were used, and 87 pieces were damaged during the operation. Of these 87 pieces, 58 were damaged when inserted into the introducer, though this kind of loss tended to decrease with training, and 29 pieces were damaged during application to the wound. Seprafilm can be used safely and efficiently during laparoscopic surgery.

Carboxymethylcellulose Sodium↗

Histopathological study of the spreading neoplastic cells in cervical glands and surface epithelia in cervical intra-epithelial neoplasia and microinvasive squamous cell carcinoma: Ki-67 immunostaining is a useful marker for pathological diagnosis from the gland involvement site.

The purpose of the present study was to clarify the spreading status of neoplastic cells in the cervical glands and surface epithelia in cervical intra-epithelial neoplasia (CIN) and microinvasive squamous cell carcinoma (MiSCC), and to evaluate the diagnostic usefulness of Ki-67 immunostaining from the gland involvement (GI) site. Cervical conization samples from 120 patients, including 110 with CIN (CIN1, n=2; CIN2, n=21; CIN3, n=87) and 10 patients with MiSCC, was examined using HE and Ki-67 immunostaining. The linear extent, lateral extent in the surface epithelia and depth of GI were significantly increased from CIN1 to MiSCC. A significant correlation was found between the linear extent and lateral extent, between the linear extent and depth, and between the lateral extent and depth. These results indicated that the size of the surface epithelial lesion and the depth in CIN gradually increased in accordance with the grade of CIN, and that GI became deeper according to the increase in the size of the surface epithelial lesion. The Ki-67 labeling index in the GI site gradually increased from CIN1 to MiSCC, which indicated that Ki-67 immunostaining is a useful marker for the pathological diagnosis of CIN from the GI site.

Adult↗

Accurate preoperative diagnosis and laparoscopic removal of the cavitated non-communicated uterine horn for obstructive Mullerian anomalies.

Obstructive Mullerian anomalies cause severe dysmenorrhea following menarche as a result of disturbed menstrual outflow. Therefore, surgical management such as extirpation of the obstructive uterine horn is required for treatment of these patients. It is necessary to have a detailed understanding of the pathological conditions of the pelvic organs and urinary system prior to surgery. We report three cases of reproductive, nulligravid patients diagnosed as having obstructive Mullerian anomalies. Preoperative accurate diagnosis was obtained by magnetic resonance imaging (MRI) and 3-D computed tomography (CT) angiography. Laparoscopic resection of the rudimentary uterine horn was performed safely and completely, and resolved all problems. MRI and 3-D CT angiography are useful tools for diagnosing complex Mullerian anomalies, and operative laparoscopy may be an alternative treatment for these cases.

Adolescent↗

Effectiveness of microlaparoscopy in the diagnosis of premature ovarian failure.

AIM: Premature ovarian failure (POF) cases contain a mixture of cases possessing follicles (type A) and those depleted of follicles (type B). Differentiation between the two types is important because the treatment policy and pregnancy prognosis vary greatly. The objective of this study was to examine the usefulness of microlaparoscopy in the differentiation of types A and B. METHODS: Among 66 patient with POF diagnosed at our department between May 1996 and April 2004, 47 patients who gave informed consent and underwent microlaparoscopy were studied. The cases were divided into four groups based on the laparoscopic ovarian morphology. These groups were analyzed with respect to patient background, blood hormone levels, antinuclear antibody positive rate, visualization of the ovary on transvaginal ultrasonography, presence or absence of follicles in biopsy specimen, and number of follicles. RESULT: No significant differences in patient background and serum hormone levels were observed between groups. There was a tendency of increase in antinuclear antibody positive rate, increase in proportion of cases with follicles, and increase in number of follicles as the ovarian morphology approached normal. Transvaginal ultrasonography failed to identify the ovary in all the patients. CONCLUSION: Microlaparoscopy is useful in the differentiation between type A and B POF, and is further expected to become an indicator of response to treatment.

Adult↗

Pathological study of Fitz-Hugh-Curtis syndrome evaluated from fallopian tube damage.

AIM: To analyze the causative pathogen, surgical indication and fallopian tube damages in Fitz-Hugh-Curtis syndrome (FHCS) cases diagnosed by laparoscopy. METHODS: Laparoscopic findings in 3568 cases at our hospital were reviewed retrospectively. Of these, 108 cases were diagnosed to have FHCS and examined for causative pathogens. Chlamydial infection was determined by both Chlamydia trachomatis (CT) antigens in the cervix and serum anti-CT antibodies. One hundred and four cases with chronic FHCS were divided into hydrosalpinx (H) and normal (N) groups based on the fallopian tube conditions, and were compared for patient profiles, tubal functions and perihepatic adhesion. RESULTS: Among surgical indications, tubal infertility and ectopic pregnancy were more often associated with FHCS than any other diseases. The primary causative pathogen of FHCS was CT. In 104 cases with chronic FHCS, there was no difference in CT antigen positivity between the H and N groups. The serum anti-CT IgG titer also did not differ between the two groups, although anti-CT IgA was slightly higher in the H group. The peritubal adhesion score was significantly higher in the H group, but it was not correlated with the degree of perihepatic adhesion. CONCLUSION: While some FHCS patients had severe fallopian tube dysfunctions, such as hydrosalpinx and tubal occlusion, 30% of FHCS cases did not show any abnormalities in the fallopian tube. Our study indicated that the severity of tubal dysfunctions associated with FHCS is determined by the host's reactivity to CT.

Adult↗

Effectiveness of intraoperative ultrasound in reducing recurrent fibroids during laparoscopic myomectomy.

OBJECTIVE: To detect and study residual fibroids, their recurrence after laparoscopic myomectomy (LM) and the risk factors. STUDY DESIGN: A prospective analysis (Canadian Task Force classification II-1) was conducted at a university-affiliated hospital. A total of 42 patients who underwent LM underwent contact ultrasonography (CUS) and transvaginal ultrasonography (TVUS) to detect residual fibroids. After LM, additional enucleation guided by CUS was performed, and then the final residual fibroids were detected using TVUS. The frequency of postoperative residual fibroids and their characteristics were evaluated. All cases were followed for > 6 months postoperatively to assess recurrence. RESULTS: The total number of fibroids enucleated in the initial enucleation was 201, or 4.8 per patient. Median diameter of the largest fibroids for each patient was 60 mm. There were 25 additional fibroids enucleated with CUS guidance. Their median diameter was 12.1 mm. After the additional enucleation, 33 final residual fibroids were identified by TVUS. Their median diameter was 9.0 mm, which was significantly smaller than those of the additionally enucleated fibroids (p = 0.002). The frequency of patients with residual fibroids was calculated in relation to the number of enucleated fibroids. The results showed that the frequency tended to increase as the number of fibroids increased and that almost all the patients (7 of 8 cases, 87.5%) with > or = 10 fibroids had residual ones. The median follow-up period was 31 months (range, 6-37), and 8 cases (19.0%) had recurrent fibroids. The recurrence rate in patients with > or = 10 fibroids was 50%. CONCLUSION: Intraoperative CUS was useful in detecting and enucleating residualfibroids. As the number of enucleated fibroids increased, the risk of residual fibroids and recurrence tended to increase.

Adult↗

Beta2-microglobulin required for cell surface expression of blastocyst MHC.

Blastocyst MHC is a mouse MHC class Ib gene that is selectively expressed in blastocysts and placenta like human HLA-G, which protect fetal trophoblasts and some tumor cells from NK cell attack, and in TAP-dependent expression on the cell surface. We expressed blastocyst MHC cDNA in beta2-deficient EL-4 S3 or beta2m-transfected EL-4 S3 cells. In parental EL-4 S3 cells, only 47-kDa blastocyst MHC protein was expressed and retained in the cytoplasm. However, additional 51-kDa blastocyst MHC protein was expressed on the surface of beta2m-transfected EL-4 S3 cells. The 51-kDa protein was resistant to Endo-H, whereas the 47-kDa protein was sensitive for Endo-H. The results suggested that beta2m as well as TAP was necessary for the transportation of blastocyst MHC from endoplasmic reticulum to cell surfaces through the Golgi apparatus, similar to other MHC class I molecules.

Animals↗