A case of recurrent type 1 diabetes mellitus with insulitis of transplanted pancreas in simultaneous pancreas-kidney transplantation from cardiac death donor.

AIMS/HYPOTHESIS: A 41-year-old woman undergoing simultaneous pancreas-kidney transplantation from an HLA-mismatched cardiac death donor abruptly developed overt hyperglycaemia under standard immunosuppressive therapy at 48 months after transplantation. Unexpectedly, we found insulitis in the transplanted pancreas and characterised the insulitis. METHODS: Pancreas graft biopsies were performed 3 years before and after the development of hyperglycaemia and the specimens were examined histologically. RESULTS: Insulitis was absent in the first biopsy, although oxidative DNA changes revealed by 8-hydroxy-2'-deoxyguanosine (8-OHdG) staining were diffusely present both in islet cells and exocrine cells. No Ki67-positive proliferating cells were seen in the islets. Anti-glutamic acid decarboxylase antibody was undetectable 6 months earlier but increased to 6.3 U/l at the development of hyperglycaemia. The level of anti-insulinoma-associated protein 2 antibody was 18.5 U/l. Insulin secretion was severely suppressed and insulin therapy was resumed. In the second biopsy, although acute allograft rejection was minimal, insulin-positive beta cells were markedly reduced, and glucagon-positive alpha cells predominated. CD3-positive T lymphocytes, CD8-positive cytotoxic T lymphocytes and CD68-positive macrophages infiltrated around and into islets. The infiltrating cells expressed Fas ligand as well as granzyme B. More than 80% of islets were affected by insulitis. 8-OHdG-positive cells were also present in islets and exocrine tissue. The percentage of Ki67-positive cells in total islet cells was 1.5%. There were no TUNEL-positive apoptotic cells in the islet cells. CONCLUSIONS/INTERPRETATION: The histological features of insulitis in transplanted pancreas were consistent with common type 1 diabetes mellitus, but the clinical course of the recurrence appeared to be more rapid.

Serum high-molecular weight adiponectin is related to early postprandial glycemic increases and gastric emptying in patients with type 2 diabetes mellitus.

BACKGROUND: Postprandial hyperglycemia and hyperlipidemia are frequently associated with type 2 diabetes mellitus. The aim of the present study is to investigate the clinical determinants of postprandial glycemia and lipemia, especially serum high-molecular weight adiponectin. METHODS: Twenty-seven diabetic patients treated with diet alone and 13 healthy volunteers took liquid test meal containing 53 g carbohydrate and 47 g lipid, dosed with nonradioactive isotope (13)C-acetate. Venous blood and breath samples were obtained for 180 min after the meal. Gastric emptying was evaluated by peak excretion time of (13)CO(2) in the breath samples. Delayed gastric emptying was defined as peak excretion time > 2.5 h (mean + 2 SD in the healthy volunteers). RESULTS: Diabetic patients showed delayed insulin secretion, postprandial hyperglycemia and hyperlipidemia compared with control. Postprandial glycemic increases significantly correlated with enhanced gastric emptying. Serum high-molecular weight adiponectin correlated with postprandial glycemic increases at 30 and 60 min after meal (r = 0.42, p < 0.05; r = 0.37, p < 0.05, respectively). Serum high-molecular weight adiponectin also correlated with gastric emptying (versus peak excretion time r = - 0.58, p < 0.05). In addition, diabetic patients with delayed gastric emptying showed the suppressed postprandial glycemia with lower serum high-molecular weight adiponectin than those with normal gastric emptying. On the other hand, postprandial increases in serum triglyceride were not related to serum high-molecular weight adiponectin or gastric emptying, but significantly related to liver function test (serum transaminases) and body mass index. CONCLUSIONS: Early postprandial glycemic increases were related to elevated serum high-molecular weight adiponectin, which might be associated with enhanced gastric emptying.

Spontaneous acute subdural haematoma and intracerebral haemorrhage in patient with HELLP syndrome: case report.

HELLP (haemolysis, elevated liver enzymes, and low platelet count) syndrome can result in a fatal intracranial haemorrhage during the perinatal period. We report treatment of a 32-year-old primigravida who fell into a deep coma during labour with fetal distress, complicated by a spontaneous acute subdural haematoma and intracerebral haemorrhage. Simultaneous emergency operations, evacuation of the acute subdural haematoma and a caesarean section, were performed, during which a diagnosis of HELLP syndrome with disseminated intravascular coagulation was made. Both mother and infant recovered, though hemiparesis persisted in the mother. Patients with HELLP syndrome should be managed as high-risk, which requires an excellent working relationship of the physicians involved. Prompt recognition of intracranial haemorrhagic complications and neurosurgical intervention are particularly important.

The retarded rate of acid-catalyzed solvolysis of glycoside bonds between reducing-end glucose residue and ceramide in glycosphingolipids compared with that of glycoside bonds between hexopyranosides.

Rates of acid-catalyzed solvolysis of glycoside bonds in glycosphingolipids were compared to establish a basis for conducting saccharide analysis. Permethylated globotetraosylceramide and asialogangliotriaosylceramide as model compounds for methylation and sugar composition analysis, respectively, were solvolyzed under acidic conditions and the sugar components thus obtained were determined at specified times by gas liquid chromatography, after they had been derivatized. Reducing-end glucose residues in both compounds were liberated more slowly than other sugar residues. Glycoside bonds between reducing-end glucose and ceramide in glycosphingolipids would thus appear to be more resistant towards acid-catalysed solvolysis than other glycoside bonds between hexopyranosides.

Cervical afferent pathway to inferior oblique motoneuron in the cat.

The neuronal pathway implicated in the vertical cervico-ocular reflex (COR) was investigated electrophysiologically in chloralose anesthetized cats. The effect of bilateral C2 dorsal root afferent stimulation on inferior oblique motoneurons (IO-MN) was investigated by intracellular recording. Control disynaptic excitatory postsynaptic potentials elicited in IO-MNs following stimulation of the contralateral anterior semicircular canal nerve (ACN) were invariably facilitated by conditioning stimulation to both ipsilateral and contralateral C2 dorsal roots (DR) in all motoneurons tested. This result indicates that inputs from the C2 DR of both sides and the contralateral ACN converge onto secondary vestibular neurons ('common interneurons') which project directly to the IO-MN. Common interneurons mediating vestibular and cervical excitation to the IO-MNs were studied in the vestibular nuclei on the side opposite to the motoneurons by extracellular recording. Nineteen vestibular neurons were identified as common interneurons; they were distributed in the caudal half of the lateral nucleus and the rostral half of the descending nucleus. The present experiment provides electrophysiological evidence of the projection of upper cervical afferents to the ipsilateral vestibular nuclei. The difference in neuronal organization between the horizontal and vertical COR is also briefly discussed.

The isolation of cultured oligodendrocytes from rat optic nerve.

Oligodendrocytes, the myelin-forming cells of the central nervous system, were cultured from newborn rat optic nerve employing a modified method of McCarthy and DeVellis, and cultured oligodendrocytes were analyzed for characteristic morphology and maturation events by phase contrast, MBP immunofluorescent and scanning electron microscopy. Two types of oligodendrocytes, type 1 OLs and type 2 OLs, on poly-L-lysine-coated dishes were obtained. Type 1 OLs showed extensive networks of processes and type 2 OLs elaborated membranous expansion along extensive networks, and it was demonstrated that type 2 OLs developed from type 1 OLs by phase contrast-microscopic observation. By scanning electron-microscopic observation, membrane structure in type 2 OLs clearly corresponded to membranous expansion. Our results suggest that type 1 OLs and type 2 OLs are in the different states of the myelinogenesis, and that membranous expansion and membrane structure in type 2 OLs correspond to myelin membrane.

The regulation of oxytocin receptor expression in human myometrial monolayer culture.

Clarification of the mechanism of oxytocin (OT)-induced contraction of the uterus seems to be essential for the elucidation of the mechanism of the initiation of labor. Although it has been suggested that estradiol (E) and progesterone (P) are involved in the expression of oxytocin receptors (OTRs), no consensus opinion has been formed on this topic. Thus we recently assessed the effects of E and P on OTR expression using cultures of human uterine cells and we examined the changes in the expressed OTRs following treatment of the cell with exogenous OT. The following results were obtained: 1. The total cellular concentration of OTRs as measured in the myometrial crude membrane fraction (total OTR concentration) showed an increase dependent on the E concentration and on the length of the incubation period in the presence of E. This increase following treatment with E (10(-7) M) was suppressed by simultaneous treatment with P in a concentration-dependent manner; being suppressed by 50% when the concentration of P was 2.7 x 10(-6) M (E/P ratio = 0.037). When 1 nM OT was added to the culture, the total OTR concentration did not change within the first 30 minutes of incubation, but decreased by about 70% twenty-four hours after addition of OT. 2. The concentration of OTRs at the cell surface (surface OTR concentration), as measured in myometrial cells that adhered to the culture plate, did not change with time when the cells were cultured at 4 degrees C in the presence of 1 nM OT. However, when cultured at 37 degrees C, the surface OTR concentration showed decreases dependent on the concentration of OT added and the time after the addition of OT. This change was observed within 60 minutes after the addition of OT to the culture. This decrease in surface OTR concentration was suppressed by concanavalin A (ConA), an inhibition of the internalization of cell surface receptors. These results indicate that in humans also, the expression of myometrial OTRs is regulated by change in the E/P ratio. The present study also revealed that OTRs, once expressed, soon disappear from the cell surface in the presence of exogenous OT (due to internalization of OTRs, i.e., dislocation of OTRs from the cell surface to the inside of the cells), and that prolonged exposure to OT even leads to the disappearance of intracellular OTRs. The present study thus suggests that the expression of human myometrial OTRs is regulated by E and P, and that an agonist-induced desensitization mechanism at the receptor level, similar to that reported for beta-adrenergic receptors, is also operating in this receptor.

Application of an Al-Ti alloy target in surface analysis by X-ray photoelectron spectroscopy.

An X-ray target of an Al-Ti alloy was used in the determination of the composition of the surface layer (0-200 A thick) of a Cu-Zn alloy.

Application of esca to semi-quantitative surface and state analysis of iron oxides.

In order to study oxidation states in the surface analysis of solids, a semi-quantitative ESCA (X-ray photoelectron spectroscopy-XPS and X-ray-excited Auger-electron spectroscopy-XAES) method was proposed. Taking iron-oxygen compounds as examples, the summation or subtraction technique was utilized in the analysis of ESCA spectra to identify the co-ordination type and crystal structure. The "XAES intensity factors" for the lines Cr L(3)M(2,3)M(4,5), Ti L(3)M(2,3)M(4,5) and from Fe L(3)M(4,5)M(4,5) to Zn L(3)M(4,5)M(4,5) were estimated and their occurrence discussed.

Analytical applications of ESCA to samples in powder form.

Analytical applications of ESCA to samples in powder form are described. These include the estimation of atomic ratios of metallic elements to oxygen in some typical oxides, the construction of calibration curves and the investigation of precipitation phenomena. The calibration curve method is useful in combination with the calculation method when the appropriate sample preparation has been carried out.

ESCA Investigation of the surface of mixed precipitates.

Quantitative ESCA has been used for studying the surface of mixed precipitates. When the precipitation is complete and homogeneous, as with titanium and zirconium hydroxides, almost complete correlation is found between the atomic ratio of the elements in the precipitate and that estimated by the Hirokawa-Ebel method, but not for incomplete mixed precipitation, as with copper and silver hydroxides. When precipitated copper and silver are reduced to the metals with iron or zinc, the surface is enriched in copper.

A case-control study on risk factors for leukemia in a district of Japan.

A case control study was performed with 142 leukemia patients and 284 controls matched for age and sex. Occupation, birth order, past medical history, and drinking and smoking habits were compared in these two groups. Persons born first or fourth were found to have a higher incidence of leukemia. History of a fracture was one of the risk factors for acute leukemia, and a history of gastroduodenal ulcer was a risk factor for chronic leukemia. This may suggest that extensive exposure to X-rays in diagnosis and treatment is a risk factor for leukemia. There was a significant dose-response relationship between the amount of smoking and the incidence of acute nonlymphocytic leukemia, but not between the amount of alcohol consumption and the incidence of leukemia. Thus, smoking was one of the risk factors for acute leukemia.

Acute anterior and posterior ischemic optic neuropathy.

Criteria for diagnosis of acute ischemic optic neuropathy (acute ION) were proposed: acute ION was divided into two groups, ie, acute anterior ischemic optic neuropathy (acute AION) and acute posterior ischemic optic neuropathy (acute PION). Acute AION is characterized by pale or sometimes hyperemic papilledema in the early stage, followed by postneuritic optic atrophy. In contrast, acute PION shows normal optic disk in the early stage followed by development of simple optic atrophy. We analyzed 31 eyes of 24 cases with acute AION and 20 eyes of 18 cases with acute PION. The diagnostic criteria and clinical findings in both types were compared. Through analyses of the etiological factors, acute PION was thought to be acute retrobulbar optic neuritis caused by ischemic changes in the optic nerve. Laboratory examinations of serum lipids, hemostatic and fibrinolytic activities in cases of acute ION did not reveal significant differences from the findings in cases with retinal vein occlusion and diabetic retinopathy. In the early stage of acute ION, steroid therapy gave rise to some improvement of the visual function but showed no effect in the late stage of the disease.

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.

Single fiber electromyography in cat extraocular muscles.

Single fiber electromyography (SFEMG) was carried out in the extraocular muscles (EOMs) of 40 adult cats, and the recordings were analyzed. The small fluctuations of the interpotential interval (IPI), so-called "jitter phenomenon", were represented quantitatively as a mean consecutive difference (MCD). The MCD and the IPI recorded from the orbital layer of the rectus muscles were similar among the 4 rectus muscles: the MCD and the mean IPI measured in 58 motor units from the 4 rectus muscles averaged 25.82 microseconds and 0.94 ms, respectively. These values were similar to those recorded from the oblique muscles. The MCD and IPI were 34.12 microseconds and 1.70 ms, respectively, in the retractor bulbi muscles and 45.54 microseconds and 1.23 ms, respectively, in the triceps muscles. The MCDs of the rectus and inferior oblique muscles were considerably smaller than those in the retractor bulbi and the skeletal muscles.

[The distribution of actin on cultured oligodendrocytes from rat optic nerve].

Two types of cultured oligodendrocytes (OLs) from rat optic nerve were analyzed for the distribution of actin, a major contractile protein, using immunocytochemistry. Type 1 OLs showed extensive network of processes and type 2 OLs showed elaborate membranous expansion along an extensive network. Actin was diffusely stained on an extensive network of processes in type 1 OLs and on the distal portion of membranous expansion in type 2 OLs, but some oligodendrocytes were not stained. It was demonstrated that the distribution of actin in type 1 OLs and type 2 OLs varied with development in oligodendrocyte differentiation and maturation. Our results suggest that undifferentiated and immature oligodendrocytes display remarkable cell movement to search for target axons, and differentiated and mature oligodendrocytes do not require cell movement when myelination has finished. In conclusion, it is considered that actin plays an important role in myelinogenesis.

[A case of suprasellar arachnoid cyst followed up for a long time].

We followed a case of suprasellar arachnoid cyst for 12 years. The patient was a sixteen-year-old girl without particular problems in her general condition. She showed optic atrophy in both eyes and optic nerve hypoplasia with an inferotemporal quandranopsia in the left eye. A suprasellar arachnoid cyst communicating with the tubarachnoid space was found to extend into the sella turcica as an empty sella. A cyst wall was resected and a cyst-peritoneal shunt performed. After 12 years from the operation, sensitivity was slightly depressed in the visual field where it had already been disturbed. Although there are few reports in the literature on involvement of the optic nerves and chiasma by suprasellar arachnoid cysts, papilledema and optic atrophy are often found in children, and infero-temporal quandranopsia or homonymous hemianopsia have been reported. Visual field defects were most likely caused by compression of the optic nerve by cyst or prolonged papilledema. We also suspect that some kind of disturbance to the optic nerve occurred during extension of the arachnoid cyst as an empty sella, or during formation of arachnoid cyst in the fetus stage.

[Responses in extraocular muscles from magnetic stimulation of the cat brain].

In 12 adult cats, pulsed magnetic stimuli were applied to the scalp, with the center of the stimulating coil over the interparietal bone. Recordings were obtained from the superior rectus, lateral rectus and inferior oblique muscles with concentric needle electrodes. In all cats, single muscle action potentials were recorded easily in all the muscles examined with latencies of 3.2-37.5 ms (mean 15.6ms). These responses disappeared after intravenous administration of pancuronium bromide. Furthermore, the elicited action potentials in the inferior oblique muscles disappeared by amputation of the inferior oblique branch of the oculomotor nerve. In some motor units, stronger stimuli shortened the latency of the responses. In most units, however, stimulus intensity did not influence the latency of responses over a wide range. The mean latency (15.6ms) is coincident with the latency of cortical spike potentials preceding voluntary saccades. This remarkable coincidence suggests that our pulsed magnetic stimuli have a very strong possibility of generation of human saccades. Thus, magnetic stimulation of the scalp may provide further information relevant to the normal operation of the oculomotor system.

[Intraperitoneal (i.p.) chemotherapy in the management of ovarian cancer--pharmacokinetics and methods of sequential i.p. chemotherapy].

We studied the systemic efficacy of intraperitoneal (i.p.) chemotherapy which has been used as a locoregional chemotherapy for ovarian cancer. The following results were obtained. 1) The CDDP-based i.v. and i.p. regimen obtained a 90% response rate for ascites and a 35% response rate for measurable lesions. 2) According to the pharmacokinetic analysis of i.p.-chemotherapy, BH-AC showed a regional advantage because of a high AUC-ratio. Meanwhile CDDP and etoposide demonstrated a low AUC-ratio, that has direct effects in the abdominal cavity and anti-tumor effects from the vascular side of the tumor. 3) We performed sequential i.p.-chemotherapy using three types of drainage tubes according to the stage and the size of the residual tumor at operation. The abdomino-centesis using a peel-away introducer for the poor risk patients was very easy and this method could carry out sequential CDDP-i.p.. No patients had signs or symptoms of chemical peritonitis. 4) On the base of these date, we have designed a protocol of i.p. chemotherapy for patients with ovarian cancer.