Glucose as regulator of glucose transport activity and glucose-transporter mRNA in hamster beta-cell line.

To investigate the role of glucose in regulating glucose transporters in pancreatic beta-cells, we studied the hamster clonal beta-cell line HIT-T15, which retains responsiveness to glucose. Northern blot analysis demonstrates that GLUT2 and GLUT1 mRNA are abundant in HIT cells. After a 24-h culture with various concentrations of glucose (0-22.2 mM [0-400 mg/dl]), the GLUT2 mRNA level in HIT cells increased by 40% at 22.2 mM (400 mg/dl) glucose compared with 11.1 mM (200 mg/dl) without a change in mRNA stability. It also decreased proportionally to the reduction of glucose concentration. Glucose deprivation resulted in a decrease of GLUT2 mRNA to an almost undetectable level, with a marked increase in the degradation rate of mRNA. In contrast, the GLUT1 mRNA was not affected by glucose. We show that glucose uptake is highest in HIT cells incubated at 2.8-5.5 mM (50-99 mg/dl) glucose for 24 h, and that levels in cells cultured at 0 mM (0 mg/dl) and 22.2 mM (400 mg/dl) glucose decrease to approximately 20% of the maximum level. This decrease is consistent with the effects of glucose on glucose-stimulated insulin secretion in HIT cells. Our results indicate that glucose is involved in regulating GLUT2 mRNA and glucose uptake activity and that the glucose responsiveness of the insulin secretion correlates with the glucose-induced change in glucose uptake activity in HIT cells.

Production of rabbit antibody specific for amino-terminal residues of cholecystokinin octapeptide (CCK-8) by selective suppression of cross-reactive antibody response.

Antibody specific for the amino-terminal region of cholecystokinin octapeptide (CCK-8) was generated in a highly reproducible way in New Zealand white rabbits by a novel immunization procedure which involves immunization with CCK-8 peptide conjugate coupled with keyhole limpet hemocyanin (KLH) and inhibiting cross-reacting antibody formation by treatment of the animals with a potent tolerogenic conjugate of beta-alanyl-tetragastrin and a copolymer of D-glutamic acid and D-lysine (D-GL). The antisera thus produced specifically react with an amino-terminal region of CCK-8 but not with the non-sulfate form of CCK-8, nor with the carboxy-terminal region which shares a cross-reactive determinant among gastrin and cholecystokinin-related peptides (caerulein, CCK-4, CCK-8, CCK-33 and CCK-39). The antisera produced by this method allowed us to measure specifically CCK in extracts from tissue such as duodenum containing gastrin and CCK at comparable levels.

Glucagon and glucagon-like immunoreactive cells in mid- and hindgut carcinoids.

The occurrence of glucagon/glucagon-like immunoreactivity in 31 small intestinal, 34 rectal and 18 appendiceal carcinoids were investigated immunocytochemically using, sequence specific antisera. Glucagon/GLI immunoreactive cells were found in five small-intestinal and five rectal carcinoids, but none were observed in any of the appendiceal carcinoids examined. Glucagon/GLI immunoreactive cells constituted a minor cell population, except in one rectal carcinoid, where most of the tumour cells were of this type. Glucagon/GLI immunoreactive cells were detected with only some sequence-specific antisera, and not with antisera directed against the rest of the glucagon/glicentin molecule. This might indicate that these cells contain a molecule which shares some antigenic binding sites with glucagon/glicentin rather than genuine glucagon/glicentin. It is concluded that this finding contributes to explain why hindgut carcinoids rarely give rise to symptoms related to neuro-endocrine product(s).

Analysis of where and which types of proteinases participate in lysosomal proteinase processing using bafilomycin A1 and Helicobacter pylori Vac A toxin.

Lysosomal proteinases are translated as preproforms, transported through the Golgi apparatus as proforms, and localized in lysosomes as mature forms. In this study, we analyzed which subclass of proteinases participates in the processing of lysosomal proteinases using Bafilomycin A1, a vacuolar ATPase inhibitor. Bafilomycin A1 raises lysosomal pH resulting in the degradation of lysosomal proteinases such as cathepsins B, D, and L. Twenty-four hours after the withdrawal of Bafilomycin A1, NIH3T3 cells possess these proteinases in amounts and activities similar to those in cells cultured in DMEM and 5% BCS. In the presence of various proteinase inhibitors, procathepsin processing is disturbed by E-64-d, resulting in abnormal processing of cathepsins D and L, but not by APMSF, Pepstatin A, or CA-074. In the presence of Helicobacter pylori Vac A toxin, which prevents vesicular transport from late endosomes to lysosomes, the processing of procathepsins B and D occurs, while that of procathepsin L does not. Thus, procathepsins B and D are converted to their mature forms in late endosomes, while procathepsin L is processed to the mature form after its arrival in lysosomes by some cysteine proteinase other than cathepsin B.

Some cellular characteristics of somatostatin neurons and terminals in the periventricular nucleus of the rat hypothalamus and median eminence. Electron microscopic immunohistochemistry.

Somatostatin neuronal perikarya and their processes, presumably dendrites, in the periventricular nucleus of the rat hypothalamus and terminals in the median eminence were observed by electron microscopic immunohistochemistry. Neuronal perikarya and processes contained immunoreactive dense granules (100-120 nm in diameter) and other cellular components such as polysomes, rER membranes occasionally showed high electron density. Few axo-somatic terminals were found on the somatostatin neurons, but we could detect a number of preterminal axons on immunoreactive processes, presumably dendrites. Therefore, we considered that somatostatin neurons receive mainly neuronal input through axo-dendritic synapses rather than through axo-somatic ones. In the somatostatin terminals in the external layer of the median eminence immunoreactivity was completely restricted on the granules.

Detection of catecholamine and luteinizing hormone-releasing hormone (LH-RH) containing nerve endings in the median eminence and the organon vasculosum laminae terminalis by fluorescence histochemistry and immunohistochemistry on the same microscopic sections.

Distribution of catecholamine (CA) and LH-RH nerve endings in the median eminence (ME) and the organon vasculosum laminae terminalis (OVLT) of the rat was investigated by application of fluorescence histochemistry and immunohistochemistry on the same sections of the tissue. In the ME, those two kinds of endings coexisted in the lateral portion of the middle part of ME, and in the wall of tuberoinfundibular sulcus, where they might be considered to have functional correlation. In the OVLT they were also distributed in fairly near distance, but they were not so closely associated as observed in the ME.

The arcuate nucleus as a primary site of satiety effect of leptin in rats.

The obese (ob) gene encodes a fat cell-derived circulating satiety factor (leptin) that is involved in the regulation of energy homeostasis. In the present study, we examined effects of i.c.v. injection of recombinant human leptin on food intake and body weight gain in rats. We also studied effects of direct microinjections of leptin into the arcuate nucleus (Arc), ventromedial hypothalamus (VMH), and lateral hypothalamus (LH). A single i.c.v. injection of recombinant human leptin (0.25-2.0 micrograms/rat) reduced significantly and dose-dependently food intake and body weight gain in rats. Microinjections (0.125-0.5 microgram/site) into the bilateral Arc, VMH, and LH caused dose-related decreases in food intake and body weight gain as compared with vehicle-treated groups with a rank order of potency; Arc > VMH = LH. The present study provides the first direct evidence that the Arc is a primary site of satiety effect of leptin.

Postnatal ontogeny of catecholamine and somatostatin neuron systems in the median eminence of the rat as revealed by a colocalization technique.

The ontogenetic development of catecholamine (CA) and somatostatin containing nerve terminals in the rostral, central and caudal median eminence (ME) of the rat was investigated by combining fluorescence histochemistry and immunohistochemistry in the same tissue section. Somatostatin terminals were detected earlier in development than CA terminals and had already appeared in the lateral part of the external layer of the central ME by the 1st postnatal day. CA nerve terminals were first observed in the same region of the ME on the 7th postnatal day. At about this stage both types of terminals seemed to show early signs of a correlation in their distribution which became progressively closer as maturation proceeded. Their distribution reached a stable condition in density and pattern on the 21st postnatal day when the majority were found in the lateral part of the external layer of the central ME, in the basal part of the brain just dorsal to the tuberoinfundibular sulcus and in the upper and lower labia of the tuberoinfundibular stalk. They subsequently matured to the adult pattern of stabilized distribution and anatomical relationship. It was also established that on the 28th postnatal day the somatostatin immunoreactive terminals reached a greater concentration than at any other stage of development including that of the adult.

An alignment method for mammographic X-ray spectroscopy under clinical conditions.

This paper describes an alignment method for mammographic X-ray spectroscopy under clinical conditions. A pinhole, a fluorescent screen, a laser device and the case for a detector are used for alignment of the focal spot, a collimator and a detector. The method determines the line between the focal spot and the point of interest in an X-ray field radiographically. The method allows alignment for both central axis and off-axis directions.

Intracranial chondroma.

Two cases of intracranial chondroma are reported. In both cases, the tumor was located extradurally in the middle cranial fossa. The tumors were removed piecemeal, but total removal proved to be impossible.

Dural sinus thrombosis in Behçet's disease--a case report.

Sinus thrombosis, which causes pseudotumor cerebri, is difficult to diagnose with conventional imaging apparatus, CT and MRI. We report a case of sinus thrombosis due to Behçet's disease diagnosed by magnetic resonance angiography (MRA). A forty-five-year old man with severe headache and ocular pain was examined. Since he had a history of oral ulcers, genital ulcers, and erythema nodosum venous thrombosis, he had been diagnosed as having Behçet's disease previously. He showed marked papilledema bilaterally but no obvious intracranial lesion was observed by CT and MRI. On MRA, an obstruction was apparent at the sagittal sinus. Anticoagulant and steroid therapy were effective to relieve the headache, and to improve the constricted visual field temporarily. By MRA, blood flow was confirmed in the sagittal sinus. However, mild papilledema persisted and finally vision was lost because of optic atrophy.

Cervical flexion myelopathy associated with cervical spina bifida occulta--case report.

A 16-year-old boy presented with segmental muscular atrophy of the bilateral distal upper extremities. Cervical spine x-ray films showed occult spina bifida from C-1 to T-1 associated with an abnormal long club-like bone located parallel to the epidural space between C-5 and C-7. In neck flexion, the cervical spinal cord was stretched and compressed to the posterior aspect of the vertebral body. Moreover, the dorsally placed abnormal bone migrated ventrally, indenting the dorsal portion of the spinal cord. This is quite an unusual case of so-called "flexion myelopathy," aggravated by the abnormal bone located dorsally.

[Prophylactic administration of an antibiotic in neurosurgical operations--penetration of cefotiam into the cerebrospinal fluid].

Various antibiotics are widely used for the purpose of protection against postoperative infections. Neurosurgeon must select the effective antibiotics to bacterium, that which penetrated enough to the intracranial organ through the blood-brain barrier. Eighteen cases with ventriculo-peritoneal shunt received intravenous drip infusion of cefotiam (CTM) and the concentration of CTM in blood and cerebrospinal fluid (CSF) was measured. The conclusion drawn from this study on penetration of CTM is summarized as follows: The concentration of CTM in CSF and its ratio to that of serum (CSF/serum %) showed the values of 0.543 microgram/ml (3.66%) in the group of 1 g CTM injection and 0.900 microgram/ml (4.02%) in the group of 2 g CTM injection, 2 hours after an intravenous administration. The concentration of CTM in CSF were gradually decreased in comparison with antibiotic levels in the blood. The most cases of 1 g or 2 g intravenous administration were able to get the sufficient concentration in the CSF, which exceeds MIC (minimal inhibitory concentration) against Gram-positive cocci and Gram-negative bacilli. As a result of penetration of CTM into cerebrospinal fluid, we recommend the intermittent intravenous injection of 2 g CTM for prophylaxis of postoperative infections.

[A study on penetration of antibiotics into the cerebrospinal fluid after the ventriculo-peritoneal shunt (author's transl)].

Neurological surgeon must select the effective antibiotics to bacterium, that which penetrated enough to the intracranial organ through the blood-brain barrier. In this study, we measured the concentration of cefotiam (CTM), cephalothin (CET), cephacetrile (CEC) and sulbenicillin (SBPC) into the cerebrospinal fluid in the non inflammatory cases with V-P shunt. 1. Antibiotic concentrations in CSF and CSF/serum ratio (%) were evaluated at 2 hours after an intravenous administration of antibiotics. CTM; 0.543 microgram/ml (3.66%), CET; 1.84 micrograms/ml (7.45%), CEC; 1.77 micrograms/ml (7.5%), SBPC, 6.15 micrograms/ml (5.58%). 2. The peak of antibiotic concentration in CSF appeared from 2 hours after administration and gradually decreased. In cellular tumor cases, the penetration of antibiotics into CSF showed similar levels as the cerebrovascular disease cases. 3. Concentration of CTM in CSF was higher than MIC of CTM to S. pyogenes and E. coli.

[A clinicopathological study of the cranial "histiocytosis X" (AUTHOR'S TRANSL)].

A clinicopathological study of 13 cases of cranial histiocytosis X was discussed. The patients were mostly infants, and those under 15 years of age numbered eight. Hematological, blood chemistrical, and other laboratorical prodedures have produced no direct diagnostic value. Radiographically, this lesion in skull appeared to be a lytic defect in all cases. Histologically, they were unified by a localized or generalized histiocytic proliferation. Eosinophilic cells were also found throughout the lesional tissue. There were some slight histological variations from case to case, for instance, in the degree of infiltration by eosinophilic cells, and the number of the histiocytic cells. However, the basic histological pattern remained the same in all cases. The lesion may already have extended into the neighboring osseous tissues. Sections of the neighboring osseous tissues showed the intertrabecular spaces to be infiltrated by the same type of lesion. The patient related trauma to the onset of symptoms in 6 cases, but it is true that we know nothing of the etiology of this lesion. The treatment of choice is the adequate surgical extirpation. It seems fair to conclude that wide surgical excision with postoperative irradiation or with chemotherapy may be expected to affect a clinical cure.

[A case of enchondroma of the skull base--a manifestation of generalized chondromatosis (author's transl)].

Cartilaginous tumors arising from the skull or intracranial structures are infrequently seen. There have been 112 such cases reported. They are most commonly found in the middle fossa and the parasellar region. They arise from cartilaginous rests in the base of the skull, particularly around the basal synchondroses. Occasional cases of intracranial growths also show lesions in peripheral bones. The authors have encountered a case in which chondroma was a manifestation of generalized chondromatosis. The patient was a 25-year-old male, who complained of diplopia in May 1975. He noted ptosis in his left eyelid which gradually progressed over a 2 months period to complete ptosis. When he was admitted in July 1975 to Nagoya National Hospital, the neurologic signs revealed a palsy of the IIIrd cranial nerve on the left side. The patient has been suffering from generalized enchondromatosis during a period of 20 years. Roentgenograms revealed extensive areas of bone destruction and pathological calcification of the phalanx, ribs, hip bone, femur, tibia, ulna and radius. Roentgenograms of the skull revealed calcification on the left middle fossa. After a left carotid angiography, an osteoplastic craniotomy was performed on August 5, 1975. The tumor was located in the extradural space of the middle cranial fossa. After incision of the capsule, gelatinous semisolid tissue was removed piecemeal. Its total removal proved to be impossible. Histological examination of this tumor revealed enchondroma.

[MRI of traumatic anosmia].

On MRI, the authors were able to demonstrate the contused lesions of bilateral rectal gyri near the crista galli in all of five cases of traumatic anosmia. Thin slice coronal and sagittal images of MRI were very useful for detecting the lesions. Conventional CT scans failed to demonstrate the lesions in 3 out of the 5 cases, but, even in these cases, MRI was able to clearly depict the contused lesions of bilateral rectal gyri. Therefore, the depicted lesions of bilateral rectal gyri on MRI can be evidence of traumatic anosmia and this may be very significant in medico-legal cases.

[2 Cases of cystic meningioma].

Although meningiomas are usually a solid and firm tumor, some are associated with diagnostically confusing large cysts. The authors experienced two cases of meningioma associated with large cyst (cystic meningioma). The first case was a 20-year-old female. She was admitted because of blurred vision. On admission she was slightly apathetic and showed bilateral papilledema. Computed tomography showed a large area of low density in both frontal regions. CT scan after intravenous contrast enhancement revealed an enhancing mural nodule attached to the falx. At operation, dark-green fluid in an amount of 160 ml from the left cyst and 50 ml from the right cyst was aspirated. The nodular tumor was found attached to the falx. The pathological examination revealed meningotheliomatous meningioma. Neoplastic meningothelial cells were found also in the cyst wall. The second case was a 21-year-old female. She was admitted because of weakness of the left lower extremity. Neurological examination revealed left hemiparesis. Deep tendon reflexes were hyperactive on the left side and there were positive left Babinski and Chaddock reflex. The computed tomography with contrast enhancement showed a large area of low density in the right parietal region with markedly enhancing mural nodule. At operation, 70 ml of xanthochromic fluid was aspirated. The tumor was well-defined, hard and attached to the dura. Histologically the tumor was a typical meningotheliomatous meningioma. The diagnostic value of computed tomography, the pathogenesis of cyst formation and the surgical treatment were discussed. The authors emphasized the important role of computed tomography in the diagnosis of cystic meningioma. Theories were reviewed for the pathogenesis of cyst formation. At operation, not only the mural nodule but also the cyst wall should be removed, because the neoplastic cells are presented in the cyst wall.

[A case of cerebral cryptococcosis, with special reference to computerized tomography findings].

A case of cryptococcosis simulating brain tumor was reviewed. A 66-year-old female was admitted to our hospital with chief complaint of vertigo, gait disturbance and dysarthria. These symptoms started about one year before admission and worsened. Vomiting and urinary incontinence appeared. Neurological examination revealed left cerebellar ataxia and dysarthria. In plain CT (computerized tomography) irregular ill-defined low density area was noted in the cerebellar vermis and bilateral cerebellar hemispheres. And slight ventricular dilatation was found. Irregular shape of ring-like enhancement corresponding to capsule and patchy or mottled enhancement inside the tumor were seen. Suboccipital craniectomy was performed and yellowish necrotic tumor with hard capsule was removed. Histological diagnosis was not neoplasm or tuberculoma. Postoperatively liver function progressively worsened. She died due to disseminated intravascular coagulation. Autopsy revealed typical liver cirrhosis without malignant change. 3.0 X 2.5 cm sized, slightly hard, yellowish lesion was found on upper part of cerebellar hemispheres. This had extremely necrotic tissue and a great number of cryptococcus neoformans were found. And other intracranial lesion was not confirmed. Finding of pulmonary cryptococcosis was not gained. Our case is very rare because of solitary cerebellar abscess and absence of meningitic episode or pulmonary cryptococcosis. There are three types of inflammation in cerebral cryptococcosis. The commonest manifestation is the meningitic type, the second mode is granulomatous lesion and the third and the least presentation is intracranial abscess formation. CT reveals various findings according to clinical stage. CT findings are those of meningitis, meningoencephalitis, granuloma and abscess. Cryptococcal granuloma or abscess often simulates brain abscess, glioma and metastatic brain tumor.(ABSTRACT TRUNCATED AT 250 WORDS)

[Immunological examination in patients with brain tumor and the usefulness of PHA skin test (author's transl)].

The usefulness of in vivo skin test reaction to phytohemagglutinin to assess cell-mediated immunity was investigated in 109 subjects, including 37 normal controls, 35 patients without brain tumor and 37 patients with brain tumor. Intradermal injection at forearm of 10 microgram of Bacto-phytohemagglutinin-P (PHA-P) produced an erythema within 24 hours. Upon PHA skin reaction the mean diameter of erythema was 25.3 mm with the normal in the control and 18.6 mm with other diseases than brain tumor, while it was 13.0 mm with brain tumor. Furthermore when mean diameter of those with longer than 15.0 mm are classified as positive in the PHA skin test, the normal in the control revealed 89.2% positive and the patients with brain tumor indicated only 37.8% positive. A positive reaction was observed in 10 (77.0%) of 13 benign brain tumor and 5 (26.3%) of 19 malignant brain tumor. A correlation was found between in vivo and in vitro responses to PHA, and between PHA and PPD skin reaction. In repeated PHA skin test at intervals of about one month, the skin reaction to PHA of patients with brain tumor varied depending on the clinical states. Other advantages of this test are no presensitization and it can be used repeatedly. The PHA skin test is a useful and simple screening test for cellular immune function of patients with brain tumor and as a simple method applicable clinically indicating accurate immuno-reactivity the PHA skin tests have been carried out during the course of treatment with an interval of about one month repeatedly.