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Biomedical subjects

T Tsutsui

Publications and source records attributed to T Tsutsui.

At least 73 records · Page 4Linked to original sources
PubMed2000

Unilateral intrauterine horn insemination of frozen semen in cats.

Frozen feline semen was prepared using two types of extenders, egg yolk Tris-fructose citric acid (EYT-FC) and egg yolk sodium citrate solution (EYC), and the semen qualities after thawing and the conception rates obtained by unilateral intrauterine horn insemination (UIUI) were investigated. Cats used in the experiment were six males and 11 females aged 2-12 years (the number of experimental cases was 17). For preparation of frozen semen, semen collected by the artificial vagina method was adjusted to I x 10(8) sperm/m/ and 7% glycerol, put in 250 microl straws, and then frozen using a cell freezer. The mean sperm motility after thawing was 30.0+/-9.7 (SE) % in the semen prepared with EYT-FC and 30.0+/-3.3% in the semen prepared with EYC. Four of seven animals were fertilized by UIUI using two straws in both extenders, and the conception rate was 57.1%. The mean ratios of number of kits to the number of ovulations in the inseminated side were 61.1+/-24.5% and 30.5+/-3.4% for EYT-FC and EYC, respectively, showing that the ratio tended to be higher in the semen prepared with EYT-FC. The above findings, comparing the two extenders for preparation of frozen feline semen, showed that EYT-FC is slightly superior to EYC. To increase conception and fertility rates, it may be important to increase the sperm count for insemination and to inseminate both uterine horns.

Animals↗
PubMed2000

Changes in plasma testosterone and testicular transferrin concentration, testicular histology and semen quality after treatment of testosterone-depot plus PMSG to 3 dogs with asthenozoospermia.

Three dogs diagnosed as having asthenozoospermia were given three intramuscular injections of 50 mg testosterone(T)-depot plus 250 IU pregnant mare serum gonadotropin (PMSG) at 2-week intervals, and their plasma T and testicular transferrin (Tf) concentrations, testicular histology, and semen quality were examined during the period of hormone therapy. Plasma T concentrations temporarily increased, and there was a slight improvement in spermatogenesis. Increased Tf concentrations suggested that Sertoli cell function in all three dogs was promoted by hormone treatment. The results showed that semen quality, especially the percentages of motile sperm and abnormal sperm, were improved between 1 and 5 weeks after the start of T-depot plus PMSG treatment.

Animals↗
PubMed2000

Plasma LH and progesterone levels before and after ovulation and observation of ovarian follicles by ultrasonographic diagnosis system in dogs.

Recently, canine frozen semen has been attracting attention for breeding purposes, and methods of judging ovulation and optimum timing for insemination have become important. As methods of predicting the canine ovulation, vaginal smear, plasma sex hormone levels and ultrasonographic diagnosis system (US) have been investigated in combination, but a standard technique has not yet been established. Therefore, we investigated a method of predicting canine ovulation in dogs by US, and by measuring plasma LH and progesterone (P) levels three times a day. Ovulation could be observed by detecting irregularly shaped ovarian follicles by US in six of 11 dogs (54.5%). In these dogs, the time between the LH peak and ovulation was 24-48 hr, 38.0 hr on average. The P level on the ovulation day was 1.88-2.81 ng/ml, 2.34 ng/ml on average. A value of 1.88 ng/ml was detected in one dog, but the other five dogs showed P levels of 2 ng/ml or higher. The P level on the day before ovulation was 0.8-1.56 ng/ml, 1.12 ng/ml on average. Assuming that ovulation occurred two days after the LH peak in the 11 experimental dogs, the P level was 2.12-4.06 ng/ml, 2.78 ng/ml on average. The period of a high LH level, not less than 10 ng/ml, continued for 12 hr around the LH peak. Based on these findings, to predict ovulation using US and LH level, it would be necessary for the tests to be performed several times a day. In contrast, it was shown that the day on which a plasma P level of 2 ng/ml or higher was detected by the test performed once a day corresponded to the ovulation day.

Animals↗
PubMed2000

Azoospermia of dogs with apoptotic germ cells and Leydig cells.

Apoptotic cell death in the testes of 4 dogs with azoospermia was examined. Blood plasma luteinizing hormone (LH), testosterone (T), and estradiol-17beta (E2) concentrations, and testicular transferrin (Tf) concentration as a marker of Sertoli cell function were measured in the 4 azoospermic dogs and in 5 normal dogs. The spermatids in 2 of the 4 azoospermic dogs and the Leydig cells in 3 of them exhibited apoptotic cell death. Mean LH, E2, and Tf concentrations in the 4 azoospermic dogs were significantly higher than in the normal dogs (P<0.01). These findings suggested that the azoospermia in all 4 dogs might has been caused by abnormal functions of Sertoli cells as well as Leydig cells.

Animals↗
PubMed2000

Effects of orvus ES paste on canine spermatozoal longevity after freezing and thawing.

Because of weak resistance of canine sperm to freezing, an applicable method of preparing canine frozen semen has not yet been established. We added various concentrations of Orvus ES Paste (OEP) to egg yolk Tris-fructose citrate, and investigated its effectiveness on survival of spermatozoa. Addition of 0.5-1.0% OEP to the extender for freezing canine semen was effective in prolonging post-thaw survival of spermatozoa.

Animals↗
PubMed2000

Effect of addition of Orvus ES paste to frozen canine semen extender on sperm acrosomes.

Using the triple-stain technique, we investigated whether sperm acrosomes in frozen canine semen were protected during freezing and thawing by addition of a surfactant, Orvus ES Paste (OEP), to the extender. Acrosomes were clearly shown to be protected by the addition of OEP to the entender when compared with those in sperm frozen without OEP addition (p<0.05).

Acrosome↗
PubMed2000

Intrauterine and intravaginal insemination with frozen canine semen using an extender consisting of orvus ES paste-supplemented egg yolk tris-fructose citrate.

Our previous report indicated that addition of Orvus ES Paste (OEP) to the extender of frozen canine semen protected acrosomes and maintained sperm motility after thawing. In this study, artificial insemination (AI) using the frozen semen was carried out. The frozen semen was prepared using egg yolk Tris-fructose citrate, and the final concentrations of glycerol and OEP were 7% (v/v) and 0.75% (v/v), respectively. AI was performed during the optimal mating period predicted from the peripheral plasma progesterone level. In intrauterine insemination (IUI), the bitches were laparotomized and 1 x 10(8) spermatozoa were infused into one of the uterine horns. In insemination of non-OEP supplemented semen, 3 x 10(8) spermatozoa were inseminated. In intravaginal insemination (IVI), 10-40 x 10(8) spermatozoa were inseminated. Conception was obtained in nine of 10 bitches (90.0%) that underwent IUI. The number of newborns was from 1 to 7 (mean 3.6 +/- 0.9). The mean ratio of the number of puppies to the number of ovulations in the inseminated uterine horn was 71.8%. The number of puppies did not exceed the number of ovulation in the inseminated uterine horn. Conception using non-OEP supplemented frozen semen was unsuccessful in all four bitches. In IVI, conception was not obtained in any of the six bitches that received insemination of 10 x 10(8) or 40 x 10(8) spermatozoa, but two of three bitches that received insemination of 20 x 10(8) spermatozoa were fertilized. It was shown that a high conception rate can be obtained by IUI using OEP-supplemented frozen canine semen. Developmenmt of a non-surgical method of IUI and a method of freezing canine sperm applicable to IVI is necessary.

Animals↗
PubMed2000

Induction of dog sperm capacitation by glycosaminoglycans and glycosaminoglycan amounts of oviductal and uterine fluids in bitches.

Ejaculated sperm collected from 12 beagle dogs were incubated in canine capacitation medium (CCM), supplemented with 5 microg/ml chondroitin sulfate A (CS), 5 microg/ml hyaluronic acid (HA), or 5 microg/ml heparin (HP) for 7 hr at 38 degrees C in a 5% CO2 in air atmosphere to investigate the effects of glycosaminoglycans (GAGs) on dog sperm capacitation. The percentages of motile sperm, hyperactivated sperm (%HY), and acrosome-reacted sperm (%AR) in all media were examined after 4 hr and 7 hr of incubation. The oviducts and uteri of 9 anestrous and 18 estrous beagle bitches were removed under halothane inhalation anesthesia to measure the total GAG amounts in oviductal and uterine fluids. The lumens of the ampulla of the oviducts, isthmus of the oviducts, and the uterine horns were each flushed with 1 ml HEPES-EDTA fluid. Total GAG amounts in the flush fluids obtained were measured with a spectrophotometer. Sperm motility (51-59%), %HY (79-86%), and %AR (31-36%) in CCM supplemented with CS, HA, or HP were significantly higher after 7 hr of incubation than when incubated in CCM without GAGs (P<0.01 or 0.05). The mean total GAG amounts in the fluids from the ampulla and isthmus of the oviducts and the uterine horns in the estrous bitches were higher than in the anestrous bitches. These results indicate that GAGs in the oviductal and uterine fluids in estrous bitches are associated with in vivo sperm capacitation.

Acrosome Reaction↗
PubMed2000

Ulnar neuropathy caused by a lipoma in Guyon's canal--case report.

A 74-year-old female presented with a 3-month history of compression neuropathy of the right ulnar nerve in Guyon's canal. Magnetic resonance imaging and ultrasonography revealed the location of the mass lesion. Surgical exploration discovered a lipoma pressing against both the ulnar nerve and the ulnar artery. The mass was extirpated. The postoperative course was uneventful with good function recovery.

Aged↗
PubMed2000

[Clinical evaluation of combined spinal-epidural anesthesia by needle-through-needle approach in 485 surgical cases].

We analyzed retrospectively the technical and clinical consequences of combined spinal-epidural anesthesia by needle-through-needle approach over the last two years. A Tuohy-type 18-gauge epidural needle (Perican; B. Braun Co.) and long pencil-pointed 27-gauge spinal needle (Whitacre; Becton-Dikinson Co.) were selected. Spinal anesthesia was administered with 0.5% tetracaine. A total of 485 anesthesia cases included 144 cases for lower abdominal, 193 cases for gynecological and 148 cases for orthopedic surgeries. The successful subarachnoid puncture with only one attempt was recorded in 89% of abdominal, 71% of gynecological and 72% of orthopedic cases. On the other hand, in three (0.6%) cases even with several attempts, the puncture was not possible. Inadvertent dural puncture and subarachnoid catheterization occurred in six (1.2%) and four (0.6%) cases, respectively. Inadequate spinal anesthesia was supplemented with epidural anesthesia in 13% of abdominal, 21% of gynecological and 7% of orthopedic cases. No serious complication occurred. We conclude that this needle-through-needle approach facilitates subarachnoid puncture with an ultra-fine spinal needle and subsequent epidural catheterization serves for supplemental and post-operative analgesia unless inappropriate subarachnoid indwelling occurs.

Adolescent↗
PubMed2000

Occlusive arterial diseases of the upper and lower extremities found in workers occupationally exposed to vibrating tools.

Hand-arm vibration syndrome (HAVS) is primarily a disorder of the fingers and hands. However, in some cases, vibration-exposed workers are observed to have also episodic blanching of the hands and feet. In latter cases, arteriographies of both the upper and lower extremities are necessary to diagnose the background arterial disorders. In this study, eight HAVS subjects with such disorders were examined by arteriography for differential diagnosis in cases of workers' accident compensation. In three HAVS cases with thromboangiitis obliterans, the arteriographic examination revealed obstructive changes in the palm and forearm as well as three below-knee lesions in the lower extremities. In five HAVS cases with arteriosclerosis obliterans, obstruction kinking or coiling, stenosis and/or tapering-off of the proper digital arteries were observed together with two below-knee lesions and three high lesions in the lower extremities. From the viewpoint of occupational health, palpation of superficial arteries of both the upper and lower extremities should be routinely performed during both pre-placement and periodic medical examinations for workers exposed to vibrating tools for early detection and/or prevention of any worsening of the background disorders.

Adult↗
PubMed1999

Exposure of macrophage-like cells to titanium particles does not affect bone resorption, but inhibits bone formation.

We examined the capacity of culture supernatants of macrophage-like cells exposed to titanium particles to influence bone formation and bone resorption, our aim being to elucidate the mechanism of implant loosening. A mouse macrophage-like cell line, J774, was exposed to titanium particles and the concentrations of prostaglandin E2, tumor necrosis factor-alpha, interleukin-1alpha, and interleukin-6 in the supernatants were measured. Titanium particles stimulated the J774 cells to release tumor necrosis factor-alpha, whereas prostaglandin E2, interleukin-1alpha and interleukin-6 concentrations remained low. The bone resorptive activity of the supernatants was measured by determining 45Ca release from cultured pre-labeled newborn mouse calvariae. The culture supernatants of J774 cells exposed to titanium particles showed no significant difference in bone resorptive activity in mouse calvariae from that of culture supernatants of J774 cells not exposed to titanium particles. The bone-forming activity of the supernatant was evaluated by determining bone nodule formation and alkaline phosphatase activity in cultured mouse calvaria cells. The bone-forming activity of the supernatants exposed to titanium particles was significantly decreased compared with the supernatants of unexposed J774 cells. This inhibition was reversed by the addition of anti-tumor necrosis factor-alpha neutralizing antibody. We conclude that tumor necrosis factor-alpha released from J774 cells exposed to titanium particles played an important role in the inhibition of bone formation rather than in the stimulation of bone resorption.

Alkaline Phosphatase↗
PubMed1999

A preliminary study of microcapsule suspension for hemolysis evaluation of artificial organs.

A microcapsule suspension, a substitute for animal blood in hemolysis tests, has been developed for evaluation of the absolute hemolytic properties of circulatory artificial organs. The microcapsule suspension was made by dispersing microcapsule slurry into an ethylene glycol sodium chloride solution. The microcapsule slurry was composed of a leuco dye solution and polyurethane membrane made by the reaction between aliphatic poly-isocyanate and polyamine by interfacial polycondensation. The microcapsule was a small particle containing dye inside. The microcapsule suspension was white; the diameter of the microcapsules was from 5 to 100 microns. The specific gravity of the suspension was 1.024, and the membrane was elastic. The fluid showed Newtonian characteristics, different from animal blood, and its viscosity was approximately 5.8 mPa.s. After the microcapsules were destroyed, the leuco dye was extracted with n-hexane from the suspension and was measured by spectroscopy after being colored with acid ethanol. Hemolysis can be regarded as a fatigue fracture of cell membranes rather than a static fracture. The destruction of microcapsules by a Potter type tissue grinder was observed at a low stroke number region and was compared to rat blood. Moreover, hemolysis tests of a commercially available centrifugal blood pump and the prototype of our centrifugal pump for mechanism checks were carried out with bovine blood. The hemolysis level of the prototype pump increased with time while the hemolysis level of the commercial blood pump did not change as much as that of the control when both pumps were tested with the microcapsule suspension. These results are similar to tests utilizing bovine blood. Therefore, hemolysis tests of circulatory artificial organs completed with microcapsule suspension are expected to provide results similar to tests with animal blood.

Animals↗
PubMed1999

Quantitative comparison of cytocidal effects of tetracyclines and fluoroquinolones on human periodontal ligament fibroblasts.

The cytocidal effects of tetracyclines and fluoroquinolones on human periodontal ligament fibroblasts (Pel cells) were studied. Pel cells were treated for 24 h with tetracycline (TC), demeclocycline (DMC), minocycline (MINO), chlortetracycline (CTC), tosufloxacin (TFLX), enoxacin (ENX), sparfloxacin (SPFX), lomefloxacin (LFLX) or ofloxacin (OFLX), and allowed to form colonies. The cytocidal effects of the antibacterial agents, as determined by a decrease in colony-forming efficiency, increased as the dose increased. However, CTC was an exception. As a quantitative measure of the cytocidal effect, LD50, the concentration which results in a 50% decrease in colony-forming efficiency relative to control cells, was extrapolated from the dose-response curves. The rank-order of cytocidal effects (LD50) was DMC [symbol: see text] MINO [symbol: see text] TC >> CTC. DMC, MINO and TC were at least 5.6-6.6 times more cytocidal than CTC. The cytocidal effects of the fluoroquinolones were in the following order: TFLX > ENX > SPFX > LFLX > OFLX. TFLX, ENX, SPFX and LFLX were 36.3, 11.4, 7.7 and 3.1 times more cytocidal than OFLX, respectively. Little cytocidal effect was observed when the cells were treated with either < or = 0.03 mM tetracyclines or < or = 0.01 mM fluoroquinolones. The results provide useful estimates concerning the relative toxicities against human periodontal ligament of antibacterial agents used to treat periodontitis.

Alkaline Phosphatase↗
PubMed1999

Targeted disruption of CDK4 delays cell cycle entry with enhanced p27(Kip1) activity.

The mechanism by which cyclin-dependent kinase 4 (CDK4) regulates cell cycle progression is not entirely clear. Cyclin D/CDK4 appears to initiate phosphorylation of retinoblastoma protein (Rb) leading to inactivation of the S-phase-inhibitory action of Rb. However, cyclin D/CDK4 has been postulated to act in a noncatalytic manner to regulate the cyclin E/CDK2-inhibitory activity of p27(Kip1) by sequestration. In this study we investigated the roles of CDK4 in cell cycle regulation by targeted disruption of the mouse CDK4 gene. CDK4(-/-) mice survived embryogenesis and showed growth retardation and reproductive dysfunction associated with hypoplastic seminiferous tubules in the testis and perturbed corpus luteum formation in the ovary. These phenotypes appear to be opposite to those of p27-deficient mice such as gigantism and gonadal hyperplasia. A majority of CDK4(-/-) mice developed diabetes mellitus by 6 weeks, associated with degeneration of pancreatic islets. Fibroblasts from CDK4(-/-) mouse embryos proliferated similarly to wild-type embryonic fibroblasts under conditions that promote continuous growth. However, quiescent CDK4(-/-) fibroblasts exhibited a substantial ( approximately 6-h) delay in S-phase entry after serum stimulation. This cell cycle perturbation by CDK4 disruption was associated with increased binding of p27 to cyclin E/CDK2 and diminished activation of CDK2 accompanied by impaired Rb phosphorylation. Importantly, fibroblasts from CDK4(-/-) p27(-/-) embryos displayed partially restored kinetics of the G(0)-S transition, indicating the significance of the sequestration of p27 by CDK4. These results suggest that at least part of CDK4's participation in the rate-limiting mechanism for the G(0)-S transition consists of controlling p27 activity.

Animals↗
PubMed1999

alpha1- and alpha2-adrenoceptors in BPH.

The dynamic obstruction of the bladder outlet secondary to benign prostatic hyperplasia (BPH), and the contractile properties of the human prostate are mediated primarily by alpha1-adrenoceptors. There are now at least three subtypes (A, B, and D) of alpha1-adrenoceptors, and recent work revealed that alpha1A-adrenoceptor and alpha1B-adrenoceptor may have a prime role for prostatic obstruction, and contraction of artery, respectively. Very recently, the presence of a low affinity alpha1-adrenoceptor for prazosin, named alpha1L, in the human BPH tissue has been determined. Because the DNA sequence of alpha1L-adrenoceptor has not yet been cloned, the alpha1L-adrenoceptor may be another form of the alpha1A-adrenoceptor, or another pharmacologically distinct alpha1-adrenoceptor which mediates the norepinephrine-induced contraction of the prostatic smooth muscle. Furthermore, the contribution of alpha1-adrenoceptors in the prostate to symptoms (not only obstructive, but irritative symptoms) which are elicited by prostatic obstruction remains to be determined.

Adrenergic alpha-Antagonists↗
PubMed1999

Function of contralateral testis after artificial unilateral cryptorchidism in dogs.

The effects of a cryptorchid testis on the contralateral testis were investigated after artificially producing unilateral cryptorchidism in 8 beagle dogs. Bilateral testicular biopsy and collection of spermatic vein blood and peripheral vein blood were performed at the time of the operation to produce the cryptorchidism and 52 weeks later. The testicular tissue was used for histological examination by light microscopy and measurement of the testicular transferrin (Tf) concentration by enzyme immunoassay. Plasma testosterone (T), estradiol-17 beta (E2), and luteinizing hormone (LH) levels were measured by radioimmunoassay. Semen was collected weekly and its quality was examined. No spermatogenesis was observed in the cryptorchid testes at 52 weeks after the operation, and the number of germ cells in the contralateral testes had decreased but the number of Sertoli cells did not change. The Tf concentration in both testes had also decreased. The mean total number of sperm between 48 and 52 weeks after the operation (194 x 10(6)) was less than half the number before the operation (510 x 10(6)). Mean spermatic vein plasma T levels (51 ng/ml) in the cryptorchid testes 52 weeks after the cryptorchid operation were significantly lower than before the operation (91 ng/ml; P < 0.05). By contrast, spermatic vein plasma E2 levels (80 pg/ml) were significantly higher than the values before the operation (51 pg/ml P < 0.05). The peripheral plasma LH levels decreased. These findings indicate that a large quantity of E2 secreted by the cryptorchid testis inhibits the endocrine and spermatogenic functions of the contralateral testis in the dog. In particular, it is assumed that dysfunction of the contralateral testis is associated with Sertoli cell dysfunction suggested by the low Tf concentration.

Animals↗