Improvement of insulin resistance following transsphenoidal surgery in patients with acromegaly: correlation with serum IGF-I levels.

AIM: The aim of this study was to determine the correlation between the growth hormone (GH)-insulin-like growth factor-I (IGF-I) axis and glucose intolerance in acromegaly during the early postoperative period. SUBJECTS AND METHODS: The study included 20 patients with acromegaly caused by GH-secreting pituitary adenoma who received transsphenoidal surgery in our hospital. Glucose tolerance was evaluated with oral glucose tolerance tests (OGTTs) performed during pre- and early postoperative periods (9 [7-18] days after surgery). Homeostasis model assessment of insulin resistance (HOMA-IR) and insulinogenic index (IGI) were calculated, and correlation analyses were performed between these values and the GH-IGF-I axis. Patients were divided according to postoperative changes of the axis, and glucose tolerance was compared between the groups. RESULTS: In preoperative OGTTs, nine patients had impaired glucose tolerance and two had diabetes mellitus patterns. Postoperatively, significant reduction was observed both in fasting plasma glucose levels (p<0.01) and in HOMA-IR (p<0.01), whereas IGI showed no significant change. HOMA-IR was significantly correlated with serum IGF-I levels both before (r=0.83, p<0.01) and after (r=0.57, p<0.01) surgery, although it was not correlated with serum GH levels. Patients who achieved more than 50% postoperative reduction in serum IGF-I levels showed significant improvement in OGTTs results (p<0.05). CONCLUSIONS: In patients with acromegaly, serum IGF-I levels, but not GH levels, were significantly correlated with insulin resistance. Early postoperative improvement of glucose tolerance is observed in patients who achieved postoperative reduction in serum IGF-I levels.

Comparison of physical properties and drug-releasing characteristics of white petrolatums.

White petrolatums of Japanese Pharmacopoeia grade and Sun white marketed as a cosmetic were characterized by measuring their physical properties and drug-releasing characteristics. White petrolatums of Japanese Pharmacopoeia grade available commercially in Japan were Perfecta, White 1S, Ultima, Snow, Snow V and Regent (Propeto). Penetrating stress, shear stress and spreading properties were measured as physical properties of the white petrolatums. The physical properties of white petrolatums varied, and Regent was the softest and the most spreadable ointment base. In vitro release test was performed using flow-through Franz diffusion cells. Fluorescein isothiocyanate and tetracycline hydrochloride were used as drug models. Their release characteristics varied among the tested white petrolatums, and Regent had the best release properties. Among the white petrolatums, with the exception of Regent, the release properties should depend on the distribution of drugs between white petrolatum and the receiver solution. Considerations of usability and characteristics of theprincipal agent are needed when choosing white petrolatums.

P-type ATPase diversity and evolution: the origins of ouabain sensitivity and subunit assembly.

Molecular aspects of the diversity of P-type ATPases are explored in this review. From the substrate specificities among different ATPase molecules, the existence of isoforms within a single class of pump becomes evident and it is now recognized as a universal phenomenon. From the phylogenetic analyses using a vast collection of the deduced amino acid sequences for the P-type ATPase subunits, it also becomes evident that the divergence of substrate-specificity occurred early in the evolution and has been conserved ever since. Further extensive analyses identify a set of novel isoforms that retain an ancestral characteristic of the Na+/K+-(H+/K+-)ATPases in invertebrates.

Churg-Strauss Syndrome with pleural involvement.

A 51-year-old Japanese man with Churg-Strauss Syndrome (CSS) diagnosed by pleural biopsy is described. He was hospitalized because of high fever and bilateral knee, elbow and shoulder joint pain. Chest roentgenogram and chest computed tomography (CT) scan revealed bilateral massive pleural effusion. Pleural biopsy revealed eosinophilic infiltration and necrotizing granulomas. He was treated with oral prednisolone and his symptoms improved. This is the first report of CSS diagnosed by pleural biopsy.

Pulmonary lymphangiomyomatosis (LAM) developing chylothorax.

We describe a case of pulmonary lymphangiomyomatosis (LAM) with chylothorax that developed in a 46-year-old Japanese woman. This patient exhibited clinical symptoms of dyspnea and chest X-ray showed right pleural effusion. Thoracocentesis demonstrated chylous effusion. Chest computed tomography (CT) scan revealed multiple cystic lesions. Subsequent thoracoscopy revealed the chylorrhea from swelled vessels on the diaphragm. The clinical diagnosis, based on histological examinations with biopsy specimens obtained by thoracoscopy, was pulmonary LAM. Although the hormone therapy was not effective, chylous effusion was improved by the pleurodesis. Pulmonary LAM developing chylothorax is rare in Japan.

Preparation and anticoagulant activity of fully O-sulphonated glycosaminoglycans.

Glycosaminoglycans including dermatan sulphate, hyaluronan, heparan sulphate and heparin were chemically modified by O-sulphonation. By altering the reaction conditions, products having a different degree of O-sulphonation could be obtained. Glycosaminoglycan derivatives were prepared having no free hydroxyl groups, with sulphoester group/disaccharide unit ratios of 4.0 for dermatan sulphate and hyaluronan, and sulphoester and sulphamide group/disaccharide unit ratios of 4.22 and 4.88 for heparan sulphate and heparin, respectively. 1H NMR spectroscopy showed that the fully O-sulphonated hyaluronan derivative had a glucuronate residue with an altered conformation. Since glycosaminiglycans and their derivatives are often used as anticoagulant/antithrombotic agents, their anti-amidolytic activities were determined. The anti-factor IIa activity of fully O-sulphonated dermatan sulphate, hyaluronan and heparan sulphate ranged from 40 to 80 units/mg, while no anti-factor Xa activity of the fully O-sulphonated glycosaminoglycans was detected. These values are lower than those reported for low-molecular-weight heparins and are consistent with the requirement of an antithrombin III pentasaccharide binding site for anti-factor Xa activity. Interestingly, the anti-factor Xa of heparin is lost by chemical O-sulphonation.

Inhibition of hyaluronidase by fully O-sulfonated glycosaminoglycans.

We report a new flow injection assay (FIA) method for determining hyaluronidase activity and the inhibitory effects of chemical fully O-sulfonated glycosaminoglycans on this enzyme. The products of enzymatic action on hyaluronidase can be detected by FIA using fluorometric detection with the fluorogenic reagent 2-cyanoacetamide. The major products derived from hyaluronan by the action of mammalian testicular hyaluronidase (a hydrolyase) were confirmed by (1)H NMR spectroscopy and capillary electrophoresis. The FIA method was next applied to the assay of hyman urinary hyaluronidase activity and the screening of hyaluronidase inhibitors. The human urinary hyaluronidase activity measured ranged from 46 to 59 turbidity reducing units/mg protein. Among the glycosaminoglycans only heparin showed hyaluronidase inhibition. Chemically O-sulfonated glycosaminoglycans showed IC(50) values of hyaluronidase inhibition that correlated with the degree of O-sulfonation. Heparin was found to inhibit hyaluronidase activity noncompetitively, while chemically O-sulfonated HA strongly inhibited hyaluronidase through both competitive and noncompetitive effects.

[Early phase II trial of oral etoposide administered for 21 consecutive days in patients with cervical or ovarian cancer. ETP 21 Study Group--Cervical-Ovarian Cancer Group].

We conducted multi-site early phase II trial or oral etoposide administered for 21 consecutive days in patients with cervical or ovarian cancer in cooperation with 19 institutes. Fifty mg/body of oral etoposide was administered daily for 21 consecutive days. Cycles were repeated every 28 days. In cervical cancer, 24 patients were enrolled and 17 of them were evaluated. The overall response rate including CR and PR was 23.5% (4/17). In ovarian cancer, 18 patients out of 21 enrolled were evaluated. The overall response rate was 16.7% (3/18). The primary toxicity observed was myelosuppression such as leukopenia, neutropenia, hemoglobin decrease and thrombocytopenia. Other adverse effects were anorexia, nausea, vomitting, fatigue, alopecia and stomatitis. From these results we concluded that oral etoposide administered for 21 consecutive days was effective against cervical cancer.

A histological study of the organic elements in the human enamel focusing on the extent of the odontoblast process.

Topographic and tomographic studies were conducted on the organic elements occluded in the enamel of premolars removed from young orthodontic patients by using light (transmitted) microscopy, confocal scanning laser microscopy (CLSM), scanning electron microscopy (SEM), transmission electron microscopy (TEM) on ultrathin sections and freeze-etching replicas, and energy dispersive spectroscopy (EDS) X-ray microscope (EDX) analysis. The present fine structure study aimed in particular to determine the fine structure of the enamel spindle and the extent of the odontoblast process. Organic elements in the ground-sectioned enamel corresponding to simple projections and enamel rods/spindles, enamel tufts and lamellae were identified by conventional light microscopy and subsequently examined by CLSM. Both light microscopy and CLSM indicated that a number of enamel spindles were measured about 50 microns in length, some 4-7 microns in thickness and were mostly confined to the cuspal summits and conformed to previous descriptions. SEM examination revealed some simple projections extending from the dentine into the enamel as well as clearly identifiable enamel spindles; the enamel spindles were structures intervening enamel prisms and showing morphological complexity by branching and convergence of the distal endings of the invading organic structure from dentinal tubules. EDX-analysis revealed that enamel tufts, lamellae, and spindles contained less phosphorus and calcium elements than enamel prisms. The enamel spindles had a higher content than tufts or lamellae, but this may be the result of contamination from surrounding enamel. Both conventional ultrathin-section and freeze-etching replica TEM evaluation of the dentino-enamel boundaries in particular suggested that simple projections and enamel rods/spindles were extensions of the odontoblast processes trapped in the enamel during early amelogenesis. In contrast, both SEM and TEM observations failed to identify dentinal tubule, peritubular (intratubular) dentine, membranous structures or lamina limitans surrounding the enamel spindles and simple projections occluded in the human enamel.

[Late phase II trial of oral etoposide administered for 21 consecutive days in patients with cervical cancer. ETP 21 Study Group--Cervical Cancer Group].

We conducted a multi-site late phase II trial of oral etoposide administered for 21 consecutive days in patients with cervical cancer in cooperation with 32 institutes. Fifty mg/body of oral etoposide was administered daily for 21 consecutive days. Treatment cycles were to be repeated at 4- to 5-week intervals. Eighty patients were enrolled and 70 patients were evaluated. The overall response rate (95% CI), including one complete response patient and 18 partial response patients, was 27.1% (19/70). The most commonly observed toxicity was myelosuppression such as leukopenia, neutropenia, hemoglobin decrease and thrombocytopenia. Other adverse effects were gastrointestinal toxicities such as anorexia, nausea, stomatitis and vomiting, as well as fatigue and alopecia. These adverse effects were well tolerated and controlled with medications. From these results we concluded oral etoposide administered for 21 consecutive days was an effective drug against cervical cancer.

A case of tracheal agenesis delivered of a patient with mosaic Turner's syndrome.

We describe a female infant with tracheal agenesis associated with severe complicated malformations including the cardiovascular system. The patient was born of a mother with mosaic Turner's syndrome at 35 weeks of gestation after premature rupture of the membranes during treatment for polyhydramnios. The patient died 2 days after birth and the autopsy disclosed tracheal agenesis and associated multiple anomalies.

[Phase II clinical study of RP56976 (docetaxel) in patients with carcinoma ovarii or carcinoma colli uteri].

An early phase II clinical study of RP56976 (docetaxel), a new semisynthetic agent, in patients with carcinoma ovarii or carcinoma colli uteri was undertaken by a cooperative study group of 23 institutes. Docetaxel was administered at an initial intravenous dose of 60 mg/m2 with dose-free intervals of 3-4 weeks, and its efficacy and safety were evaluated. Of the 47 patients with carcinoma ovarii enrolled, 44 patients were eligible and 36 patients completed the scheduled course of treatment. Of the 23 patients with carcinoma colli uteri enrolled, 20 patients were eligible and 15 patients completed the scheduled course of treatment. For antitumor efficacy in patients with carcinoma ovarii, 1 patient showed partial response (PR), 10 showed no changes (NC) (2 showed minor response (MR)), and 25 had progressive disease (PD). The overall response rate was 2.8% (1/36). Of patients with carcinoma colli uteri, 7 patients showed no changes (NC) (1 patient showed minor response (MR)), 8 patients had progressive disease (PD). Major adverse reactions included 64/65 (98.5%) leukopenia, 56/59 (94.9%) neutropenia, 40/60 (61.5%) decrease of hemoglobin, 12/64 (18.8%) thrombocytopenia, 30/65 (46.2%) anorexia, 23/65 (35.4%) nausea/vomiting, 37/65 (56.9%) alopecia, and 26/65 (40.0%) fatigue, all of which were mild.

Scanning electron microscopy of elastic system fibers in the articular disc of the rat mandibular joint.

Elastic system fibers (ESFs), i.e., microfibrils (putative oxytalan fibers), elaunin and elastic (true elastin) fibers, in the rat mandibular joint were studied mainly using scanning electron microscopy (SEM) and light microscopy (LM) with the aid of image processing. The present quantitative analysis using LM showed that the articular disc and capsule, which are the sites that receive physical compressive force during mastication, contained more ESFs than the articular cartilage of the mandibular joint. In addition, oxytalan fibers were the principal ESFs in all the articular components (capsule, articular disc, supraossous layer of articular surfaces and articular cartilage). Subsequently, ESFs in the articular disc, which contained more thick ESFs, were closely examined by SEM using both collagen- and elastin-digestion methods. SEM showed networks of microfibrils beneath the articular surfaces (superior and inferior layers) in the thin central portion of the articular disc; the principal microfibrils ran at nearly right angles to the collagen fibers. The microfibrils were cemented with amorphous elastin, thickened and shifted towards interconnecting oblique fibers and many main ESF trunks, which were oriented in the direction of the layered wavy collagen fibers and parallel to the direction of applied force, to sustain the mechanical force. From the superior and inferior layers, the main ESFs shifted towards the middle portion of the disc, transitional zone (synovial osteochondral junction) and the other articular components, showing no specific directivity. Transmission mission electron microscopy revealed that the thick main ESFs in the elastic network were elaunin fibers. The present study indicated that ESFs unite, branch and therefore construct an extensive and complicated protective stretchable network, which is interposed with the less tensible collagen network in the mandibular articular disc.

Histochemical studies of mitochondrial activities of cultured corneal endothelial cells of cat during wound-healing.

The corneal endothelial cells of the cat were cultured, and in 3 weeks the cells became confluent and formed a cell sheet. The cells were stained with rhodamine 123 and nitroblue tetrazolium (NBT). The frequencies of cells stained by these two methods changed during the 3-week culture, but at the end of the culture the frequencies were stabilized at 58 +/- 3.9 (mean +/- SD) % for the rhodamine 123 staining method and 35 +/- 3.7% for the NBT method. An oval wound of 600 X 400 microns was made in the center of the cultured endothelial cell sheet, and the cell migration was observed by phase contrast microscopy. Six hours after the wounding, the cells began to migrate toward the center of the wounded area, and in 24 hours the denuded area was almost completely covered by migrated cells. In 48 hours the wound was tightly covered by the migrated cells. One hour after the wounding, the cells in the wound margin were strongly stained by rhodamine 123, but not by NBT. The cultured cell sheet was divided into 4 concentric areas, ie, S1 was the center of the wound, S2 was the 100-microns-wide zone inside the original wound edge, S3 was the 100-microns-wide zone outside the original wound edge, and S4 was the 100-microns-wide zone outside the S3 zone. Topographical changes in rhodamine- and NBT-stained cells were observed over a 48-hour period. Rhodamine-positive cells increased in the S1 area, while a decrease occurred in the S2 and S3 areas. NBT-positive cells peaked at 12 hours after the wounding and decreased thereafter. It was concluded that the endothelial cells migrating to cover the wounded area showed a significant enhancement of mitochondrial activity, as indicated by the rhodamine 123 staining, but the succinic dehydrogenase activity revealed by NBT staining was rather suppressed.

Monoclonal antibody RG1 against rat Müller cells recognizes optic nerve and cerebral cortex glia in culture.

A monoclonal antibody designated as RG1 was produced by the fusion of mouse myeloma cells with spleen cells from mice immunized with rat retinas. The monoclonal antibody was shown to bind to Müller cells of the rat retina by the avidin-biotin peroxidase complex method. The screening of antibody-producing hybridomas and the analysis of the monoclonal antibody obtained were performed by enzyme-linked immunosorbent assay. The optic nerve, cerebral cortex and dorsal root ganglia were cultured from 17-day embryonic rats and maintained for 17-20 days. Expressions of the RG1 antigen were examined by indirect immunofluorescence in the cultured glial cells. In both the optic nerve and the cerebral cortex glial cells the RG1 antigen was expressed; however, the RG1 antigen was not expressed in cultured Schwann cells throughout the whole culture period. Our results suggest that the RG1 antigen is a common antigen in the supporting neuroectodermal glial cells in the central nervous system.

Cell movements in a living mammalian tissue: long-term observation of individual cells in wounded corneal endothelia of cats.

Although the cells in tissues are known to be motile under special conditions (e.g., during tissue turnover or wound healing), there are not many reports that polygonal cells covering an area without leaving any gaps are also capable of movement. In the present study, cell movements (cell shifting and rearrangement) in a living mammalian eye tissue were documented by identifying and locating individual cells over intervals as long as 100 days. Cat corneal endothelium, a monolayered cell sheet, was wounded by removing a small number (about 180) of endothelial cells from the internal lining of the cornea. Healing of the wounded tissue was observed with a wide-view specular microscope applied to the outer surface of the cornea, enabling us to identify individual cells for as long as two to three months. Cells surrounding the wound underwent areal enlargement, elongated toward the wound, and shifted to cover the wound surface. During days 4-7, cells became rearranged by changing neighbors in such a way that they retained their enlarged size but recovered their non-elongated, original shape. This pattern of cell rearrangement was interpreted by a computer simulation which assumed that cells shorten their boundary length while maintaining contacts with contiguous cells. After day 7, the enlarged cells adjacent to the wounded area gradually contracted and pulled surrounding cells toward the wounded area. These movements were followed by a temporary halt in cell shifting, then by a recovery of shifting and cell elongation. These movements are interpreted as a result of the contractility of endothelial cell microfilaments.

A case of mesenchymal chondrosarcoma of the orbit.

A case of mesenchymal chondrosarcoma occurring in the right orbit of a 84-year-old Japanese female was reported. The ultrastructural findings of tumour are composed of three cell types: well-differentiated cartilaginous cells, undifferentiated cells and transitional cells. The well-differentiated cells showed scalloped cytoplasmic membranes, numerous mitochondria showing dense electron matrix, various size of lipid granules, and abundant amount of extracellular matrix. The extracellular matrix were collagen fibres and ground substances and matrix vesicle. The undifferentiated cells showed smooth cytoplasmic membranes, large nuclei resembling primitive mesenchymal cells.