Thermal and endocrine regulation of gonadal apoptosis during sex differentiation in pejerrey Odontesthes bonariensis.

The fate of the differentiating gonads in pejerrey Odontesthes bonariensis is determined by the environmental water temperature experienced by the larvae during the critical period of sex determination. We previously reported a link between apoptosis, temperature and sex differentiation in this species. To clarify this link, we subjected larvae to thermal and endocrine treatments between hatching and the onset of histological sex differentiation of the gonads and assessed the patterns of gonadal development and apoptosis by light microscopic histology and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) analysis, respectively. Apoptotic labeling was widespread among somatic cells of the anterior region of the right gonads of fish reared at the male-producing temperature (MPT) and part of the fish at the mixed sex-producing temperature prior to sex differentiation. In contrast, TUNEL-positive cells were rarely observed in gonads at the female-producing temperature (FPT). Administration of exogenous estrogen completely prevented MPT-induced masculinization, induced feminization and reduced gonadal apoptosis, whereas an aromatase inhibitor (fadrozole) induced TUNEL signals in the gonads of FPT-reared larvae. These results provide strong evidence that apoptosis in somatic cells in the right lobe of the gonads might play a key role in testicular differentiation in pejerrey and that estrogens are involved in the regulation of this process.

[Induction chemoradiation followed by resection through anterior approach for superior sulcus tumor].

Two cases of induction chemoradiation followed by surgical resection were reported. A 53-year-old man and a 68-year-old man who had been suffering form alleviate pain in their left shoulder and arms were referred to our hospital. Physical examination revealed Horner's syndrome on the left side in both patients. A transcutaneous needle biopsy confirmed non-small-cell lung cancer. Under the diagnosis of superior sulcus tumor in stage IIIB (T4N0M0), induction chemotherapy and radiation were given. After tumor reduction, they underwent resection through cervical anterior approach because subclavian vessel invasion was suspected. The clavicle was divided for the resection and reconstruction of subclavian artery in case 2. For the treatment of anterior superior sulcus tumors, anterior approach provides a safe and effective exposure.

Low-volume muscle endurance training prevents decrease in muscle oxidative and endurance function during 21-day forearm immobilization.

AIM: To examine the effects of low-volume muscle endurance training on muscle oxidative capacity, endurance and strength of the forearm muscle during 21-day forearm immobilization (IMM-21d). METHODS: The non-dominant arm (n = 15) was immobilized for 21 days with a cast and assigned to an immobilization-only group (Imm-group; n = 7) or an immobilization with training group (Imm+Tr-group; n = 8). Training comprised dynamic handgrip exercise at 30% of pre-intervention maximal voluntary contraction (MVC) at 1 Hz until exhaustion, twice a week during the immobilization period. The duration of each exercise session was 51.7 +/- 3.4 s (mean +/- SE). Muscle oxidative capacity was evaluated by the time constant for phosphocreatine recovery (tau(off)PCr) after a submaximal handgrip exercise using (31)phosphorus-magnetic resonance spectroscopy. An endurance test was performed at 30% of pre-intervention MVC, at 1 Hz, until exhaustion. RESULTS: tau(off)PCr was significantly prolonged in the Imm-group after 21 days (42.0 +/- 2.8 and 64.2 +/- 5.1 s, pre- and post-intervention respectively; P < 0.01) but did not change for the Imm+Tr-group (50.3 +/- 3.0 and 48.8 +/- 5.0 s, ns). Endurance decreased significantly for the Imm-group (55.1 +/- 5.1 and 44.7 +/- 4.6 s, P < 0.05) but did not change for the Imm+Tr-group (47.9 +/- 3.0 and 51.7 +/- 4.0 s, ns). MVC decreased similarly in both groups (P < 0.01). CONCLUSIONS: Twice-weekly muscle endurance training sessions, each lasting approx. 50 s, effectively prevented a decrease in muscle oxidative capacity and endurance; however, there was no effect on MVC decline with IMM-21d.

Malignant hyperthermia in a patient with Graves' disease during subtotal thyroidectomy.

We report the case of a 31-year-old man with Graves' disease who manifested malignant hyperthermia during subtotal thyroidectomy. His past medical history and family history were unremarkable. Before surgery, his condition was well controlled with propylthiouracil, beta-adrenergic blocker and iodine. During the operation, anesthesia was induced by intravenous injection of vecuronium and thiopental, followed by suxamethonium for endotracheal intubation. Anesthesia was maintained with nitrous oxide and sevoflurane. One hour after induction of anesthesia, his end tidal carbon dioxide concentration (ET(CO2)) increased from 40 to 50 mmHg, heart rate increased from 90 to 100 beats per min and body temperature began to rise at a rate of 0.3 degrees C per 15 min. Suspecting thyroid storm, propranolol 0.4 mg and methylprednisolone 1,500 mg were administered, which, however, had little effect. Despite the lack of muscular rigidity, the diagnosis of malignant hyperthermia was made based on respiratory acidosis. Sevoflurane was discontinued and dantrolene was given by intravenous bolus. Soon after the treatment, ET(CO2), heart rate and body temperature started to fall to normal levels. His laboratory findings showed abnormally elevated serum creatine phosphokinase and myoglobin but normal thyroid hormone levels. Since dantrolene is efficacious in thyrotoxic crisis and malignant hyperthermia, an immediate intravenous administration of dantrolene should be considered when a hypermetabolic state occurs during anesthesia in surgical treatment for a patient with Graves' disease.

Absorption in the visible region of YSZ implanted with Ag ions.

Ag ions were implanted into YSZ (yttrium-stabilized (cubic) zirconia) single crystals in two different energy regimes: kiloelectron volt and megaelectron volt. Optical absorption spectra were measured in the visible region at each stage in the annealing process of the sample. Depth profiles of Ag for the samples implanted at the energy of 20 keV were measured by X-ray photoelectron spectroscopy (XPS). For the samples implanted with Ag at the low energy of 20 keV, one large absorption peak appeared in the wavelength ranging from 470 to 536 nm, depending on the dose of Ag ions. As the sample was heated to 1000 degrees C, the intensity of the absorption peak decreased gradually, but a small, broad peak remains even at the temperature of 1000 degrees C. For the samples implanted with 2.8x10(16) Ag ion cm(-2) at the high energy of 3 MeV, one broad absorption peak was observed at around 470 nm. As the sample was heated sequentially to high temperatures, the peak gradually decreased and almost disappeared at 400 degrees C. When the sample was further heated to even higher temperatures, the absorption peak at 514 nm reappeared at 1000 degrees C and grew with heating time.

Facial nerve repair accomplished by the interposition of a collagen nerve guide.

OBJECT: Facial nerve paralysis due to a surgical procedure or trauma is a frequently observed complication. The authors evaluated facial nerve repair achieved by the interposition of a collagen nerve guide. METHODS: Ten cats were divided into three groups. Group 1 consisted of six animals in which a 5-mm facial nerve segment on one side was resected and replaced by a collagen tube that was sutured to bridge both nerve stumps. On the opposite side a 5-mm segment of facial nerve was resected, reversed 180 degrees, and sutured to the stumps as an autograft nerve. Group 2 consisted of two cats in which the collagen nerve guide was interposed on one side and the nerve on the other side was left intact. Group 3 consisted of two cats in which a reversed autograft nerve was placed on one side and the nerve on the other side was left intact. Histological, electrophysiological, and horseradish peroxidase labeling examinations were performed starting 3 weeks after surgery. Light and electron microscopic examinations of collagen tube-implanted specimens revealed a well-vascularized regenerated nerve. The electrophysiological study confirmed the recovery of electrical activity in regenerated axons. Horseradish peroxidase labeling also confirmed restoration of the whole facial nerve tract. CONCLUSIONS: The collagen nerve guide shows great promise as a nerve conduit.

Skin flap storage with intracellular and extracellular solutions containing trehalose.

To evaluate and compare the effects of IT-Kyoto (IT-K) solution (intracellular type) and ET-Kyoto (ET-K) solution (extracellular type) on long-term skin flap preservation (from 24-120 hours), Japanese white rabbits were used as an animal model and Euro-Collins (EC) solution was used as a comparative solution. A total of 180 animals were divided randomly into 15 groups (N = 12) for 24, 48, 72, 96, and 120 hours of flap storage in three solutions. The survival rate, weight change, and histological appearance of flaps were compared among the three solutions. The survival rates in the IT-K and ET-K solutions were the same: 100%, 100%, 100%, 92%, and 50% in flaps preserved for 24, 48, 72, 96, and 120 hours, which were higher (p < 0.01 in the 72-, 96-, and 120-hour groups) than those in the EC solution group: 100%, 75%, 33%, 8.3%, and 0%. The flap weight gain in the IT-K and ET-K solution groups was less than that in EC solution group (p < 0.01). Histologically, flaps in the IT-K and ET-K solution groups showed a better appearance than those in the EC solution group. It was concluded that in a rabbit model IT-K and ET-K solutions are superior to EC solution for long-term skin flap preservation, and the safe preservation period for IT-K and ET-K solutions is approximately 96 hours. It was also concluded that the effects of intracellular (IT-K)-type solution and extracellular (ET-K)-type solution on rabbit skin flap preservation are similar.

Facial nerve repair using a collagen conduit in cats.

We evaluated facial nerve regeneration using a collagen tube as a nerve conduit in five cats. In three 5 mm of the facial nerve were resected, a collagen tube was implanted, and a 5 mm segment of the opposite facial nerve was resected, reversed 180 degrees, and sutured back as an autologous nerve graft. In one a collagen tube was implanted on one side, and in the remaining one a 5 mm nerve segment was reversed. Histological, electrophysiological, and horseradish peroxidase labelling examinations were carried out 4-24 weeks postoperatively. Histological study showed that the nerve was well vascularised and regenerated. Electrophysiological examination confirmed the recovery of evoked electromyograms through to the regenerated axons. Horseradish peroxidase examination also confirmed restoration of the whole facial nerve. The collagen tube is an efficient nerve conduit.

A large Japanese family with Machado-Joseph disease: clinical and genetic analysis.

We report clinical and genetic studies on a large Japanese family with Machado-Joseph disease (MJD), in which various different clinical phenotypes were seen in the same family, i.e., cerebellar ataxia type, severe amyotrophy type, and young-onset parkinsonism type. In addition, patients with very mild symptoms (formes frustes) were encountered. The expansion of the CAG repeat at the MJD locus ranged from 64 to 71 in 7 affected and 4 presymptomatic individuals. In our family, no clear inverse correlation was noted between the length of CAG-expansion and the age of onset, or the clinical phenotypes. Hyporeflexia was a common manifestation seen in 5 patients. It has been reported that the presence of peripheral neuropathy in MJD is associated with smaller increase in the CAG repeats; findings in our family conform with this observation.

Alterations in the enzyme activity and protein contents of protein disulfide isomerase in rat tissues during fasting and refeeding.

Protein disulfide isomerase (PDI) is an enzyme that participates in the formation of disulfide bonds. It is also known to be the subunits of some enzymes and the membrane-associated thyroid hormone-binding protein. In this study, we measured the quantitative distribution of PDI protein in rat tissues and examined the relationship between protein level and enzyme activity in PDI during fasting and refeeding. Western blotting with specific anti-PDI antiserum detected the PDI protein band of 55 kd. Among several tissues, liver contained the largest amount of PDI protein, followed by kidney and fat, in which one-third to one-fourth of the hepatic PDI protein existed. The PDI protein band was also detected in heart and muscle. Fasting for 3 days decreased PDI protein levels in rat liver by 40%; control levels were recovered after 3 days of refeeding. The same change was observed in kidney. PDI activity, measured by the scrambled ribonuclease method, did not show the parallel alteration to PDI protein level in liver and kidney. Isomerase activity decreased to 50% of control values during fasting, but did not recover by refeeding. Thyroidal status did not affect either PDI protein level or isomerase activity. These findings show that fasting and refeeding affect PDI protein and enzyme activity, and that PDI protein level does not always reflect PDI activity.

Difference in dominant negative activities between mutant thyroid hormone receptors alpha1 and beta1 with an identical truncation in the extreme carboxyl-terminal tau4 domain.

Although different expression patterns of thyroid hormone receptor (TR) alpha1 and beta1 have been reported, no essential distinction has been established in their functions. Unlike the TR beta gene, a mutation in the TR alpha1 gene has never been found in patients with resistance to thyroid hormone (RTH). Previously we found a mutant TR beta with an 11-carboxyl (C)-terminal amino acid truncation (betaF451X) in a girl with severe RTH. BetaF451X is a natural mutant with disruption of the transactivation domain, tau4, and it had very strong dominant negative activities. Based on the fact that the 46 amino acid sequence in the extreme C-terminal region is identical in TR alpha1 and TR beta, except for a C-terminal three amino acid extension of TR alpha1, we constructed a mutant TR alpha1 (alphaF397X) with the identical C-terminal truncation to betaF451X, to study functional differences between TR alpha1 and beta1. Both betaF451X and alphaF397X had negligible T3 binding and transcriptional activities even with 1 microM T3. The dominant negative activities of the mutant TRs were remarkable and T3 response element (TRE)-dependent. Co-expression of betaF451X decreased the CAT activity of either wild-type TR alpha1 or beta1 at 100 nM T3 by approximately 90% on the TRE-pal2 and 70% on DR4. AlphaF397X inhibited the transcriptional activities of both wild-type TR alpha1 and beta1 by approximately 50% on TRE-pal2 and by 60% on DR4. The dominant negative potency of betaF451X was significantly stronger than that of alphaF397X on the TRE-pal2, -DR4 and chicken lysozyme silencer F2, but similar on TRE-myosin heavy chain alpha and malic enzyme. No partiality for the TR subtypes was found in the dominant negative effects of betaF451X and alphaF397X. Co-expression with RXR enhanced the dominant negative effects of alphaF397X, but not of betaF451X. The results indicate that there are different dominant negative properties between alphaF397X and betaF451X, which are TRE-dependent, despite their identical C-terminal truncation. Deletion in the tau4 domain might affect the receptor structures of TR alpha1 and beta1 differently.

Evaluation of new improved solution containing trehalose in free skin flap storage.

BACKGROUND DATA: In this study, we evaluated a new intracellular type (IT-K) solution containing trehalose in a rabbit free skin flap storage model. Trehalose is a nonreducing disaccharide that can stabilise cell membranes under various stressful conditions. MATERIAL AND METHOD: Seventy-two free skin flaps of the ear of rabbits were preserved in Euro-Collins (EC) solution or in IT-K solution for 24, 48, and 72 h at 4 degrees C. After completion of preservation, these flaps were replanted to the other ear by microsurgical techniques. Viability study and photo documentation were performed daily for 7 days. Tissue specimens were taken 24 h after vascular anastomosis, fixed in 10% formaldehyde and stained with haematoxylin and eosin (HE). Survival rates were analysed by Fisher's exact test for comparison of the two experimental groups. Values of P < 0.05 were considered to be statistically significant. RESULTS: After 7 days, a survival rate of 100% of flaps were observed in both solutions after 24 h of preservation. After preservation for 48 h in IT-K solution the survival rate was 100%. However, in EC solution survival decreased to 75% (9 of 12 preserved flaps survived). This difference increased to 33.3% (4 of 12 flaps) in EC solution and 91.6% (11 of 12 flaps) (P < 0.01) in IT-K solution when the flaps were stored for 72 h. Light microscopic examination also showed less damage in flaps preserved in IT-K solution than in these preserved in EC solution. CONCLUSION: IT-K solution was superior to EC solution in the preservation of free skin flaps on rabbit ears when stored for 48 and 72 h.

Prolongation of ischaemic tolerance of rat skeletal muscle by trehalose.

Trehalose is a disaccharide that can stabilise cell membranes under various stressful conditions. We have studied the effect of trehalose on the ischaemic tolerance of fat skeletal muscle flaps. A rat gracilis model was used to evaluate the prolongation of ischaemic preservation of the modified solution in which trehalose was substituted for glucose in Euro-Collins solution. Four groups of animals were studied. In two groups, flaps were preserved in Euro-Collins solution or trehalose Euro-Collins solution for various ischaemic intervals (6, 12, 24, 48, 72, and 96 hours). In two additional groups, after storage for various periods in the two solutions, muscle flaps were replanted into the original animals with microvascular anastomoses, and were investigated by light and electron microscopy on the seventh postoperative day. Trehalose Euro-Collins solution significantly prolonged ischaemic tolerance compared with Euro-Collins solution alone at 24 and 48 hours.

High incidence of positive autoantibodies against thyroid peroxidase and thyroglobulin in patients with sarcoidosis.

OBJECTIVE: Although abnormalities of the humoral immune system, such as increased immunoglobulin production, are known in sarcoidosis, the relationship between sarcoidosis and autoimmune disorders in uncertain. We studied the incidence of thyroid autoantibodies and the prevalence of Hashimoto's thyroiditis in patients with sarcoidosis. PATIENTS AND MEASUREMENTS: Sixty-two patients with pulmonary sarcoidosis, diagnosed by a combination of clinical, radiographic and histological findings were studied. As controls, three groups of subjects aged 40 and over without a known history of thyroid disease (60 patients with pulmonary diseases other than sarcoidosis, 88 hospital employees and 82 company workers), were also analysed. Antibodies against thyroid peroxidase (TPO-Ab) and purified thyroglobulin (Tg-Ab) were measured by radioimmunoassay and antibodies against microsomal antigen (MCHA) and thyroglobulin (TGHA), by haemagglutination. RESULTS: Seventeen of 62 patients (27.4%) had either positive TPO-Ab or Tg-Ab or both. All the patients with positive thyroid autoantibodies were of middle or advanced age, and the incidence of positive TPO-Ab/Tg-Ab in patients with sarcoidosis aged 40 and over was 54.5% in males, 32.4% in females and 37.8% overall. The prevalence was significantly higher in males compared to age-matched control males (0-7.7% in the controls), and in female patients was twice that found in controls (11.8-16.3%). Seven patients had Hashimoto's thyroiditis, indicating that the prevalence was 11-3%, and much higher than that previously reported. CONCLUSIONS: The data show a remarkably high incidence of thyroid autoantibodies in patients of middle of advanced age with sarcoidosis, especially in males, and a higher prevalence of Hashimoto's thyroiditis than in previous reports.

Preservative effects of trehalose on rat muscle flaps.

In this experiment, the effect of trehalose on the preservation of rat gracilis muscle flaps was studied. The muscle flaps were preserved in 4 degrees C Euro-Collins solution or 4 degrees C trehalose-Euro-Collins solution for 6, 12, 24, 48, 72, and 96 hours, and then replanted in the same rats using microsurgical techniques. Immediately after preservation and 1 week after replantation, the levels of adenosine triphosphate and creatine phosphate in flaps preserved in the trehalose-Euro-Collins solution were significantly higher than those in flaps preserved in the Euro-Collins solution. In the 4 degrees C Euro-Collins solution group 1 week after replantation, focal necrosis, hyaline degeneration, and proliferation of fibroblasts were seen after 12 hours of preservation, and, after 48 hours of preservation, there were no surviving flaps. In the 4 degrees C trehalose-Euro-Collins group, focal necrosis, hyaline degeneration, and proliferation of fibroblasts were seen only after 24 hours of preservation, and, after 96 hours of preservation, two of eight flaps survived. These findings indicate that trehalose was effective in the preservation of muscle flaps.

Introducing a point mutation identified in a patient with pituitary resistance to thyroid hormone (Arg 338 to Trp) into other mutant thyroid hormone receptors weakens their dominant negative activities.

Clinical resistance to thyroid hormone (RTH) has been classified into generalized resistance to thyroid hormone (GRTH) and pituitary resistance to thyroid hormone (PRTH) types. Since similar mutations have been identified in tri-iodothyronine (T3) receptor (TR) beta gene in GRTH and PRTH, and since considerable overlap has been seen in the clinical manifestations in patients with GRTH and PRTH, two subtypes of RTH are now considered to be a continuous spectrum with the same genetic defect. A point mutation at amino acid Arg 338 to Trp (R338W) which we identified in a patient with PRTH is very interesting, since R338W has been found in several other patients with PRTH, raising the possibility that this mutation may tend to associate with a phenotype of PRTH. In our previous study, we found that R338W had relatively less impaired transcriptional potency, weaker dominant negative activity on various T3 response elements and poor homodimer formation, as compared with another GRTH mutant. In this study, to investigate the functional properties of R338W further, especially in terms of the relation between transcriptional activity and dimer formations, we introduced the R338W mutation into the mutant receptors, K443E and F451X, constructing the double mutants, R338W/K443E and R338W/ F451X. Both R338W/K443E and R338W/F451X showed negligible T3 binding and transcriptional activities. The dominant negative activities of K443E and F451X were, however, significantly weakened by introducing the R338W mutation. As a control, a double mutant G345R/K443E was constructed by introducing a point mutation, G345R, located in the same exon 9 as R338W, into the K443E mutant. Dominant negative activity did not differ between G345R/K443E and K443E. Homodimer formation was significantly reduced in the double mutants containing R338W, but not G345R. In summary, introducing the R338W mutation, but not G345R, into the mutant TR significantly weakened the dominant negative activity, despite further impairment of the T3 binding and transcriptional activities.

Preservation of skin free-flap using trehalose.

In this study, we looked for a solution able to preserve traumatically amputated tissues such as nose tip, ear, and scalp for at least 48 hr that was easy to handle and low in price. Trehalose is a nonreducing disaccharide that can protect many prokaryotes, fungi, and yeasts against adverse conditions by stabilizing cell membranes. In order to study its protective effect, 60 skin free-flaps of the ears of rabbits were preserved in Euro-Collins (EC) solution or in modified Euro-Collins solution in which 7% trehalose replaced glucose (EC + 7%T) for 24, 48, and 72 hr at 4 degrees C. After completion of preservation, these flaps were transferred to the other ear with microvascular anastomosis. After 7 days, we obtained a survival rate of 100% of flaps after preservation for 24 hr in both solutions. But, after preservation for 48 hr or longer, this decreased to 60% (6 of 10 preserved flaps survived) in EC solution and 90% (9 of 10 flaps) in EC + 7%T solution. This difference became clearer after 72 hr of preservation, when the survival rate was higher and light microscopic observation showed less damage for flaps preserved in EC + 7%T solution (20% in EC and 70% in EC + 7%T) (P < 0.05). Thus, EC + 7%T solution was superior to EC solution in the preservation of skin free-flaps of the ear of rabbits. Trehalose shows great promise in the preservation of many organs, including storage of traumatically amputated tissues such as nose tip, ear, and scalp.

[Studies on aspoxicillin for its transfer to the skin].

To evaluate the usefulness of aspoxicillin (ASPC) in the field of plastic and reconstructive surgery, we examined its transfer to the skin. 1. After intravenous drip infusion of ASPC for 1 hour at a dose of 2 g in 13 adults and at 1 g in 2 children, the mean serum ASPC concentration 1 hour after termination of the infusion was 70.46 +/- 28.05 micrograms/ml. The mean concentration in the skin tissue 1 hour after infusion in 15 patients was 32.45 +/- 18.47 micrograms/g. The rate of transfer to the skin 1 hour after infusion in the 15 patients was 52.9 +/- 29.7%. 2. The ASPC concentrations in skin tissues and the rates of its transfer to the skin did not differ significantly between 5 patients with facial surgery and 10 with surgery in the trunk or limbs. 3. To prevent postoperative infections, ASPC was intravenously drip infused twice daily for 2 approximately 3 days after operation at a dose of 2 g in adults and 1 g in children. No postoperative infection occurred in any patient, suggesting the effectiveness of this drug. In addition, no side effects or abnormalities in clinical examination values were observed.

Leukoencephalopathy induced by tegafur: serial studies of somatosensory evoked potentials and cerebrospinal fluid.

A case of leukoencephalopathy induced by tegafur, an antineoplastic derivative of 5-FU, is reported. The patient received 600 mg of tegafur p.o. for 16 days before excision of rectal cancer. After the operation, gait disturbance and mental abnormalities appeared. He became akinetic and mute within a few days following readministration of tegafur. Serial studies of brain CT, somatosensory evoked potentials (SEP) were made, and myelin basic proteins (MBP) in the cerebrospinal fluid were measured. The level of MBP was about twice the normal value and the central conduction time (CCT) of SEP was prolonged at admission. The value of MBP and CCT improved with recovery from akinetic mutism.