Long-term proliferation of mouse primordial germ cells in culture. Primordial germ cells (PGCs) are first identifiable as a population of about eight alkaline phosphatase-positive cells in the 7.0 days postcoitum mouse embryo. During the next 6 days of development they proliferate to give rise to the 25,000 cells that will establish the meiotic population. Steel factor is required for PGC survival both in vivo and in vitro and together with leukaemia inhibitory factor stimulates PGC proliferation in vitro. In feeder-dependent culture, PGCs will proliferate for up to 7 days, but their numbers eventually decline and their proliferative capacity is only a fraction of that seen in vivo. Here we report a further factor that stimulates PGC proliferation in vitro, basic fibroblast growth factor (bFGF). Furthermore, bFGF, in the presence of steel factor and leukaemia inhibitory factor, stimulates long-term proliferation of PGCs, leading to the derivation of large colonies of cells. These embryonic germ cells resemble embryonic stem cells, pluripotent cells derived from preimplantation embryos, or feeder-dependent embryonal carcinoma cells, pluripotent stem cells of PGC-derived tumours (teratomas and teratocarcinomas). To our knowledge, these results provide the first system for long-term culture of PGCs.


Naloxone-reversible effect of spantide on the spinally mediated behavioural response induced by neurokinin-2 and -3 receptor agonists. [D-Arg1, D-Trp7,9, Leu11]-substance P (spantide) was tested for antagonism against the licking, biting and scratching response induced by various neurokinin (NK) receptor agonists and bombesin (Bom) in mice. When co-administered with substance P (SP) intrathecally, spantide reduced the SP-induced behavioural responses in a dose-dependent manner. The duration of this antagonistic effect was approximately 30 min. Behavioural responses induced by physalaemin (Phy), [pGlu6, L-Pro9]-SP (6-11) (septide), [pGlu6, D-Pro7]-SP (6-11) (D-septide) and eledoisin (Ele) were also dose-dependently decreased by relatively small doses of spantide. Higher doses of spantide were needed to reduce the behavioural responses induced by [Sar9, Met (O2)11]-SP, neurokinin A (NK A) and neurokinin B (NK B). No significant effect of spantide was observed against the behavioural responses elicited by Bom. Pretreatment with naloxone, an opioid antagonist, resulted in a reversible effect on the behavioural reduction of NK-2 and NK-3 receptor agonists produced by spantide. However, the effect of spantide on the NK-1 receptor agonist-induced response was unchanged by naloxone. In homogenates of mouse spinal cord, competition studies confirmed that the binding of the opioid ligand [3H]naloxone was displaced by spantide with a low but measurable affinity. These results suggest that the behavioural response to NK-2 and NK-3 receptor agonists may be partially inhibited by spantide through the activation of opioid system in the mouse spinal cord.


Role of cyclic AMP in the release of noradrenaline from isolated rat atria. Effect of pretreatment with clenbuterol. The possible role of cyclic AMP (cAMP) on tritium overflow evoked by stimulation of the cardio-accelerant nerves was studied in rat atria preincubated with [3H]-noradrenaline. Addition of the activator of adenylate cyclase forskolin (1 mumol/l), or of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX, 100 mumol/l), did not affect both basal and evoked overflow. However, in the presence of the alpha 2-adrenoceptor antagonist yohimbine (0.03 mumol/l) both forskolin and IBMX increased the stimulation-induced transmitter overflow by 49% and 141%, respectively (compared to yohimbine 0.03 mumol/l). Thus, in rat atria the cAMP-dependent facilitation of noradrenaline release is only present when the autoinhibition exerted by activation of prejunctional alpha 2-adrenoceptors is blocked. Propranolol (0.1 mumol/l) that did not produce any effect on noradrenaline release markedly reduced the facilitatory response induced by forskolin in the presence of yohimbine. When rats were pretreated with the beta 2-adrenoceptor agonist clenbuterol (0.3 mg.kg-1, s.c., twice daily, 14 days), a treatment which desensitizes beta-adrenoceptor-mediated facilitation of noradrenaline release (Kazanietz and Enero 1989), the facilitatory effect of forskolin and IBMX in the presence of yohimbine was abolished. The results indicate that in rat atria the effect of forskolin and IBMX on noradrenaline release are only to be observed after blockade of presynaptic alpha 2-adrenoceptor autoinhibition. beta-adrenoceptor blockade or clenbuterol pretreatment decreases the facilitatory response to forskolin and hence prejunctional beta-adrenoceptor-mediated enhancement of noradrenaline release is linked to the stimulation of adenylate cyclase.


CP-96,345, a non-peptide antagonist of substance P: I. Effects on the actions mediated by substance P and related tachykinins on the guinea-pig ileum and rabbit jejunum. The effects of a non-peptide antagonist of substance P, CP-96,345, were investigated, in vitro, on the guinea-pig ileum and the rabbit jejunum. Contractions of the guinea-pig ileum, induced by substance P and neurokinin A, were specifically inhibited by the racemate (+/-)CP-96,345 (pIC50 7.8 and 7.3, respectively). The inhibition by (+/-)CP-96,345 of contractions evoked by neurokinin B and by bradykinin (pIC50 6.1 and 4.9, respectively) was attributed to unspecific effects of the antagonist. The inhibition of substance P-induced contractions of the rabbit jejunum required a 10 times higher concentration of (+/-)CP-96,345 (pIC50 = 6.8) than was required with the guinea-pig ileum. The plateau phase of contraction of the guinea-pig ileum induced by high concentrations of substance P, neurokinin A or neurokinin B, which is known to be mediated through tachykinin receptors on intrinsic cholinergic neurones, was inhibited by 200 nM (+/-)CP-96,345 but not by the inactive enantiomer, CP-96,344. This indicates a specific inhibition of these neuronal tachykinin receptors by (+/-)CP-96,345. Contractions known to be mediated by the release of substance P, such as those evoked by capsaicin and by mesenteric nerve or field stimulation, were partially inhibited by (+/-)CP-96,345 at concentrations of 200 to 600 nM. Unspecific inhibitory effects of CP-96,345, in concentrations of 1 microM or higher, were observed on histamine-induced contractions, and on the cholinergic twitch response to electrical stimulation, of the guinea-pig ileum.(ABSTRACT TRUNCATED AT 250 WORDS)


CP-96,345, a non-peptide antagonist of substance P: II. Actions on substance P-induced hypotension and bronchoconstriction, and on depressor reflexes in mammals. In order to extend the in vivo pharmacological profile of CP-96,345 ((2S,3S)-cis-2-(diphenylmethyl)-N-((2-methoxyphenyl)- methyl)-1-azabicyclo[2.2.2]octan-3-amine dihydrochloride), a non-peptide antagonist of substance P, the compound was tested against substance P-induced effects on blood pressure in rabbits and on respiration pressure in guinea-pigs. In addition, CP-96,345, and its inactive (2R,3R)-enantiomer, CP-96,344, were also tested in 2 models involving presumably substance P-mediated reflexes in the rat. The fall in blood pressure evoked by i.v. injections of substance P in the rabbit was inhibited by 0.3 mumol kg-1 CP-96,345 i.v. for at least 30 min, and almost abolished, for at least 2 h, by 3 mumol kg-1. In guinea-pigs, the bronchoconstriction induced by i.v. injections of substance P, but not that in response to histamine or bradykinin, was inhibited by CP-96,345 (0.6 and 6.0 mumol kg-1, i.v.) in a dose-dependent manner and this inhibition lasted for 40 min and 70 min, respectively. CP-96,345 (3-20 mumol kg-1, i.v.) reduced depressor reflexes in response to stimulation of peripheral capsaicin-sensitive neurones in the rat. However, the inhibition was not dose-dependent and was also seen with the inactive enantiomer, CP-96,344. This effect can hardly be attributed to receptor-specific effects of CP-96,345 and may be related to unspecific actions such as those involved in the cardiac depression exhibited by both enantiomers. The present results show that CP-96,345 is a potent antagonist of substance P in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)


CP-96,345, a non-peptide antagonist of substance P. III. Cardiovascular effects in mammals unrelated to actions on substance P receptors. The cardiovascular effects of CP-96,345, a non-peptide antagonist of substance P, were analyzed in vivo and in vitro. In the anaesthetized rat, the i.v. injection of 3 mumol kg-1 of CP-96,345 induced a fall in mean arterial blood pressure and a reduction in heart rate. Similar effects were obtained with the enantiomer CP-96,344 (2R,3R)-cis-isomer of CP-96,345) which does not interact with substance P receptors. Both enantiomers, at a concentration of 10 microM, decreased the beating frequency of the isolated atria and of the isolated perfused heart of the guinea-pig to a similar extent, and caused transient coronary dilatation. CP-96,345 (10 microM) decreased the spontaneous sinus rate, prolonged the atrioventricular-nodal conduction interval and the His-bundle conduction interval of the perfused guinea-pig heart. The intraventricular spread of conduction was markedly inhibited. During programmed stimulation 10 min after the beginning of the drug application, the effective refractory periods evaluated by stimulation with premature beats, as well as rate dependent effective refractory periods, of the atrioventricular node, of the atrial and of the ventricular myocardium, were prolonged. Sinus node recovery time was also prolonged. It was concluded that these cardiac effects of CP-96,345 were not caused by an action of the compound on substance P receptors.


Effects of the novel calcium channel blocker, anipamil, on the isolated rabbit heart. Comparison with verapamil and gallopamil. The calcium channel blocking activity of the novel phenylalkylamine derivative, anipamil, was tested on the isolated rabbit heart, in comparison with verapamil and gallopamil. Anipamil and the other calcium channel blockers lower left ventricular pressure in the same concentration range (10(-8)-10(-4) mol/l). The negative inotropic effect of anipamil is only partially reversed (nearly 65%) by rising calcium concentration in the perfusion fluid, whilst a complete recovery is observed for verapamil and gallopamil. The negative inotropic effect of anipamil is of rapid onset but long lasting, being still present 12 h after washout. On the contrary, that of gallopamil or verapamil completely disappears within 3 h of washout. Verapamil and gallopamil (10(-8)-10(-4) mol/l) depress spontaneous heart rate up to asystolia and abolish the vasopressin- and Bay K 8644-induced coronary spasm. Anipamil, on the contrary, does not modify coronary spasm elicited by both stimulants and spontaneous heart rate up to 10(-4) mol/l. These observations suggest that anipamil, in the isolated rabbit heart, possesses a peculiar pharmacological profile, since its calcium channel blocking activity is confined to the myocardial muscle.


[Gram staining and bacterial culture of stomach contents of the full-term neonate in prolonged status of ruptured membranes is not useful]. In an effort to evaluate the management of full-term neonates born after prolonged rupture of the membranes, we studied 159 full-term newborn infants born 24 hours or more after rupture of the membranes retrospectively. Gram stain and bacterial culture of a gastric aspirate of the neonates were significantly related; however, the predictive value of the Gram stain was low, both for a positive test (46%) and for a negative test (70%). A significant relation was not found between the Gram stain (presence of bacteria or greater than or equal to 4 leucocytes per high power field) or the bacterial culture on the one hand and the occurrence of clinical symptoms of infection on the other. In all cases the decision to treat the newborn with antibiotics was made on the basis of the clinical symptoms alone. In our opinion the routine study of Gram stain and bacterial culture of a gastric aspirate of full-term neonates born after prolonged rupture of the membranes is not useful; observation alone is sufficient.


Hepatitis C.

Hepatitis C. Until recently the diagnosis of non-A, non-B hepatitis was made by excluding other detectable viral infections of the liver. Progress in molecular biology made it possible to develop assays which can trace antibodies against the hepatitis C virus. This virus plays a major role in the pathogenesis of transfusion-related and sporadic non-A, non-B hepatitis and possibly of other liver diseases. Although the genome of a few isolates of the hepatitis C virus has already been decoded, the viral particles have not yet been visualized.


Degradation of myelin basic protein by a membrane-associated metalloprotease: neural distribution of the enzyme. A metalloprotease activity associated with myelin membrane preparations degrades myelin basic protein (MBP), generating a characteristic fragment designated peptide C (MBP 74-170). Using an immunoblotting assay, peptide C-generating activity was detected in mammalian, avian, reptilian, and amphibian brains. The activity was present in rat brain as early as postnatal day 1 and also in adult rat peripheral nerve. Immunohistochemistry with a monoclonal antibody to the purified enzyme revealed that the metalloprotease was present in oligodendrocytes of optic nerve, of both white and grey matter of spinal cord, and also in the cytoplasm of both myelinating and nonmyelinating Schwann cells of peripheral nerve.


Molecular mimicry and the autoimmune response to the peripheral nerve myelin P0 glycoprotein. In the Lewis rat immunisation with the myelin P0 glycoprotein can induce an inflammatory demyelinating disease of the peripheral nervous system, experimental allergic neuritis (EAN), which has many clinical and histopathological parallels with the human disease the Guillain-Barre syndrome. In view of the reported association of GBS with a number of infectious agents we have investigated whether "molecular mimicry" may occur between microbial antigens and the P0 protein that could possibly trigger a similar pathogenic autoimmune response in man. A computer search of the available protein sequence data bases identified several absolute sequence homologies between P0 and viral proteins that involve five or more consecutive amino acid residues. Four of these sequence homologies involved viral pathogens previously associated with the Guillain-Barre syndrome, namely Epstein-Barr virus (EBV), cytomegalovirus (CMV), Varicella zoster virus (VZV) and human immunodeficiency virus I (HIV I). Although, sequence homologies were also found between viral peptides and the neuritogenic determinants of P0, residues 56-71 and 180-199, these homologies proved incapable of eliciting EAN in the Lewis rat. These observations are discussed with reference to the role that molecular mimicry between T cell epitopes on pathogen derived antigens and the P0 protein may play in the pathogenesis of the Guillain-Barre syndrome.


Rapid morphological changes in bovine brain microvascular endothelial cells on extracellular matrices. The morphological changes in endothelial cells derived from bovine brain microvasculature, carotid artery, and aorta during growth on extracellular matrices were compared. All cells formed tubular structures on a basement membrane. Ultrastructural studies showed that the tubular structures had lumens surrounded by many endothelial cells. On type I collagen gel, brain microvascular endothelial cells still formed tubular structures, but the other two cell types formed confluent monolayers. However, when a second layer of collagen gel was laid over these cells, tubular structures developed within 2-3 days. Brain microvascular endothelial cells form tubular structures more readily than endothelial cells derived from large vessels on both basement membrane and type I collagen gel.


Morphometrical analysis of GFAP-positive cells in human astrocytomas. The morphometrical characteristics of human astrocytoma were examined using the shape, size, and intercellular distances of glial fibrillary acidic protein-positive cells in samples from 21 patients with various astrocytomas and normal samples resected during operations on seven patients with other brain tumors. The results showed that astrocytoma cells were rounder than normal astrocytes with fewer processes. Low-grade astrocytomas were smaller than astrocytes or rapid regrowth astrocytomas. Morphological analysis of astrocytoma may be valuable for evaluating the potential migration to adjacent tissue.


Immunohistochemical study of a newly defined oncofetal antigen (HGR-Ag) in tumors of the central nervous system. The oncofetal antigen (HGR-Ag) in brain tumors was newly defined using the monoclonal antibody derived from tissue extracts of an anaplastic astrocytoma. The HGR-Ag was purified through sequential chromatography using diethylaminoethyl-Sephadex A-50 and Con-A Sepharose affinity columns. The latter method indicated that HGR-Ag is not a glycoprotein, and Western blot analysis indicated a molecular weight of 80-90 kd. Purified antigen was used to produce antibody. This new antibody, designated H1H2, was shown to be an immunoglobulin G1 by immunodiffusion assay. Histological/immunohistochemical studies using the H1H2 antibody on paraffin and frozen tissue sections showed HGR-Ag to be intracellular rather than membrane-associated. The immunoreactivity of H1H2 was highest in malignant astrocytomas, glioblastomas, and normal fetal brain tissue. Neurinomas and certain carcinomas of other organs showed lesser, variable reactivity to H1H2. The strength of the antigen-antibody binding was correlated with the degree of malignancy in human gliomas. H1H2 bound to fetal brain tissue and undifferentiated neural tumors, but not to normal adult brain tissue, so HGR-Ag is probably a fetal oncoantigen.


Postauricular response in motor paresis with intracranial lesions. The clinical usefulness of the postauricular response (PAR) in the evaluation of motor paresis was studied in 105 patients with intracranial lesions and 25 normal volunteers. Click stimuli at 90 dB hearing level were delivered in each ear, and PARs recorded from bilateral posterior auricular muscles with reference to the vertex (Cz). No volunteer demonstrated PAR in the relaxed posture, but in tensed postures increasing muscle tone PARs were elicited with a latency of 11.5 +/- 0.7 msec and an amplitude of 6.2 +/- 2.8 muV. Sixty-two patients (59%) had high-amplitude PARs in the relaxed posture. PARs were enhanced more frequently in paretic patients than in non-paretic patients (p less than 0.01). In subcortical lesions, the PAR latency had a significant correlation with the degree of motor paresis (p less than 0.005). In patients, the enhanced PAR amplitude suggested the presence of motor paresis associated with deep-seated mass lesions. The enhanced response is thought to be associated with dysfunction of the pyramidal and extrapyramidal tracts.


Stereotactic radiosurgery using a linear accelerator. A basic and clinical study of radiosurgery using the linear accelerator (Linac) system for unremovable deep-seated brain tumors is reported. A Komai stereotactic ring was used to locate the target coordinates. The patient was laid on the Linac treatment table and held in the head fixation system. Irradiation was given in five positions. The dose profile by film dosimetry and Rando phantom was satisfactory. Seventeen tumors in 14 patients were treated. Clinical or histological diagnoses were nine metastases, one benign and two malignant gliomas, one meningioma, and one craniopharyngioma. Tumor sizes were between 8 and 30 mm. Doses were between 12 and 30 Gy. Computed tomographic evaluation after 3 months of 12 tumors in 11 survivors showed one complete remission, three partial remission, six no change, and two partial deterioration. For progressive tumors, Linac radiosurgery results are excellent.


Angiographically occult arteriovenous malformation in the septal region--case report. An occult arteriovenous malformation (AVM) in the septal region occurred in a 14-year-old boy, manifesting as headache and vomiting. Computed tomography showed a high-density mass in the septal region, faintly enhanced postcontrast. Mild hydrocephalus was also seen. Angiography revealed no abnormalities other than hydrocephalic signs. The lesion was totally removed by the transventricular approach after corticotomy of the left frontal lobe. The histological diagnosis was AVM. He was discharged without neurological or endocrinological deficits.


Foramen magnum meningioma and arachnoid cyst coinciding in the lateral cerebellomedullary cistern--case report. An extremely rare foramen magnum meningioma associated with an arachnoid cyst in the lateral cerebellomedullary cistern occurred in a 65-year-old female presenting with dizziness. Neuroimaging revealed a meningioma at the left lateral edge of the foramen magnum and an arachnoid cyst mainly located in the right lateral cerebellomedullary cistern, compressing the medulla oblongata bilaterally. After fenestration of the cyst wall and tumor removal, the clinical symptoms ameliorated. We recommend that where a foramen magnum tumor coexists with an arachnoid cyst of the posterior fossa, the tumor should be removed after shrinking the cyst to obviate the need for brainstem retraction.


Enlargement of cerebellar arteriovenous malformation associated with fenestration of the vertebral artery--case report. We report a 30-year-old male case of an enlarged cerebellar arteriovenous malformation (AVM) associated with fenestration of the vertebral artery. Hemodynamic stress resulting from fenestration of the feeding system of AVM was probably an important factor in the enlargement of the small cerebellar AVM.


Transpetrosal extreme lateral suboccipital approach for extensive posterior fossa epidermoid cyst--case report. A new transpetrosal extreme lateral suboccipital approach was adopted to totally remove an extensive posterior fossa epidermoid cyst in a 36-year-old male. The pathological behavior of intracranial epidermoid cysts may impose surgical problems during removal of the tumor. However, planning based on neuroimaging allows optimum access to tumors and microsurgery achieves safer and more complete removal.


Lectin cytochemistry combined with silver colloid staining of argyrophilic nucleolar organizer regions in human gliomas. Investigation of lectin cytochemical staining of inflammatory cells in human gliomas showed that Allomyrina dichotoma (Allo-A) cytochemistry can reliably distinguish inflammatory from neoplastic cells. Allo-A cytochemistry combined with silver colloid staining of argyrophilic nucleolar organizer regions (Ag-NORs) was performed in human gliomas. Inflammatory cells possessed usually one but sometimes two Ag-NORs and macrophages often possessed several Ag-NORs. The mean Ag-NOR number per nucleus of inflammatory cells ranged from 1.81 to 2.34, and that of neoplastic cells ranged from 2.57 to 3.53 and from 2.84 to 4.46 in low- and high-grade gliomas, respectively. The mean Ag-NOR number per nucleus of inflammatory cells was significantly smaller than that of neoplastic cells (p less than 0.001). Combined Allo-A cytochemical and silver colloid Ag-NOR staining can provide a reliable Ag-NOR number in human gliomas by distinguishing inflammatory cells.


Indications for surgery and prognosis in patients with cerebral cavernous angiomas. Seventy-three cerebral cavernous angiomas were removed microsurgically from a series of 71 patients between August, 1983 and December, 1989. This retrospective investigation assessed the current indications for surgery and determined the prognosis for patients with cerebral cavernous angioma. There were 38 males and 33 females with a mean age of 37 years. Analysis included clinical presentation and history, neuroradiological findings, indications for surgery, and postoperative course. After an average follow-up period of 15 months, 35 patients were symptom-free, 16 had improved preoperative complaints, six were unchanged, and eight had deteriorated. Microsurgical extirpation of the malformation is indicated in all symptomatic patients where neuroimaging demonstrates the presence of a readily accessible cerebral cavernoma. Surgery is recommended in cases with deep-seated lesions causing massive hemorrhage, repetitive minor bleeding, or significant long-standing and progressive neurological disabilities. Clinically silent cavernomas located in eloquent regions of the brain contraindicate surgery, but should be closely monitored. Patients presenting with convulsions or neurological deficits caused by easily accessible cavernomas of the hemispheres have the best prognosis and a negligible risk for surgical complications. Those with deep-seated lesions of eloquent regions of the brain that have bled or caused sustained neurological disorders face the highest risk for morbidity owing to the surgical intervention, requiring careful preoperative evaluation.


Risk factors for brain abscess in patients with congenital cyanotic heart disease. Brain abscess is a serious complication of congenital cyanotic heart disease. We retrospectively evaluated the risk factors for brain abscess in 21 such patients treated between 1975 and 1990 in comparison with a control group. The mean arterial oxygen saturation, arterial partial pressure of O2, arterial blood oxygen content, and base excess in patients with brain abscess were significantly lower than in the control patients. The mean arterial partial pressure of CO2, pH, hematocrit, hemoglobin, and red blood cell content in patients with brain abscess were not significantly different. Patients with congenital cyanotic heart disease may develop minute encephalomalacia due to severe hypoxemia and increased blood viscosity resulting from compensatory polycythemia. The increased blood viscosity and reduced blood flow in the microcirculation may induce cerebral thrombosis or exaggerate minute encephalomalacia during dehydration or cardiac dysfunction, and shunted blood containing infectious organisms at such sites may be followed by focal cerebritis.


Evaluation of therapeutically induced hypertension in patients with delayed cerebral vasospasm by xenon-enhanced computed tomography. Serial cerebral blood flow (CBF) measurements were made with stable xenon-enhanced computed tomography in 20 patients with angiographically confirmed ruptured intracranial aneurysms, before and during induced hypertension with continuous infusion of dopamine. All patients showed angiographic vasospasm during their course. Twelve patients without symptomatic vasospasm (Group 1) had the lowest hemispheric CBF on the craniotomy side of 31.6 +/- 6.8 ml/100 gm/min on days 4-9 (control value, 40.1 +/- 2.0 ml/100 gm/min), while the other eight patients with symptomatic vasospasm (Group 2) had the lowest hemispheric CBF on the craniotomy side of 25.0 +/- 7.6 ml/100 gm/min on days 10-14. The critical hemispheric CBF inducing neurological deficits was about 20 ml/100 gm/min in Group 2. Dysautoregulation was usually present in Groups 1 and 2, but therapeutically induced hypertension could reverse the delayed neurological deficits, if begun early at the stage of delayed vasospasm.


Treatment of unclippable giant posterior cerebral artery aneurysms with detachable balloons--report of three cases. Three patients presented with rare giant posterior cerebral artery aneurysms, clinically manifesting as cerebral ischemia, mass effect, and subarachnoid hemorrhage. All aneurysms were partially thrombosed, originated at the P2 segment, and possessed broad necks. Surgical neck clipping was difficult but proximal occlusion of the parent artery was feasible. Aneurysm occlusion sparing the parent artery was attempted in all cases, but failed because the detachable balloon did not successfully block the aneurysmal neck. All patients tolerated test occlusion at the P2 segment, so the parent artery was occluded proximally with detachable balloons, leaving the important perforating arteries unaffected. Two transient ischemic attacks were associated with the procedure. Where surgical treatment is unusually difficult, and proximal ligation or trapping just feasible, embolization with detachable balloons is an acceptable substitute.


Chronic subdural hematoma following bypass surgery--report of three cases. Bypass surgery is a safe procedure with low mortality and morbidity, and few reported surgical complications. Three patients developed postoperative chronic subdural hematoma (CSDH): two with stroke after superficial temporal artery-middle cerebral artery (STA-MCA) anastomosis and one with moyamoya disease after STA-MCA anastomosis combined with encephalomyosynangiosis. The factors inducing CSDH after revascularization in the seven reported and present cases included postoperative subdural effusion associated with brain atrophy, and postoperative anticoagulant therapy such as aspirin. CSDH may occur in patients with pre-existing brain atrophy and postoperative subdural effusion. Anticoagulant therapy should be avoided at the early postoperative stage after bypass surgery.


Bilateral posterior fossa epidural hematoma--report of two cases. Two cases of posttraumatic bilateral posterior fossa epidural hematoma are presented. Such hematomas are extremely rare, but can be surgically cured. Computed tomography helps to establish the diagnosis and determine the neurosurgical treatment.


Giant aneurysm of the azygos anterior cerebral artery associated with acute subdural hematoma--case report. A ruptured giant aneurysm of the azygos anterior cerebral artery (ACA) associated with an acute subdural hematoma (SDH) occurred in a 67-year-old male with two episodes of sudden severe headache and transient loss of consciousness. Neurologically, he had mild weakness of the left lower extremity. Computed tomography showed an elliptical heterogeneous hyperdense mass in the interhemispheric fissure in front of the corpus callosum and an acute SDH on the right. Angiography disclosed a giant aneurysm (2.8 x 2.0 cm) at the distal end of the azygos ACA. Removal of the SDH and aneurysmal neck clipping achieved a good outcome. Successive small bleedings may allow the aneurysmal dome to develop adhesions to the arachnoid membrane, and the final rupture will occur into the subdural space, resulting in a SDH.


Dynamic magnetic resonance appearance of the vertebral dissecting aneurysm--case report. A dissecting aneurysm of the vertebral artery occurred in a 38-year-old male presenting with ischemic attacks. Vertebral angiography revealed irregular narrowing and dilatation of the right vertebral artery, and retention of the contrast medium. Routine T1-weighted magnetic resonance (MR) imaging demonstrated signal voids compatible with a patent but narrowed vertebral lumen surrounded by three intensity components at the abnormality location identified on the vertebral angiograms. Dynamic MR imaging demonstrated strong enhancement of these components on the early images. These findings were consistent with the false lumen of the dissecting aneurysm. The different signal intensities were thought to reflect the blood flow rate.


Suprasellar malignant teratoma with 11-year survival--case report. Long-term survival of malignant teratoma is rare. A 16-year-old female with a suprasellar malignant teratoma survived for 11 years before succumbing to embryonal carcinoma. A suprasellar immature teratoma with increased serum alpha-fetoprotein level was verified histologically. Radiation therapy and combined chemotherapy were given after partial removal. Eleven years later, the primary lesion recurred as embryonal carcinoma.


A study of evoked potentials in the 18q-syndrome which includes the absence of the gene locus for myelin basic protein. We report evoked potential findings in a patient with 18q-syndrome (18q22.3----qter). The deletion included the locus for myelin basic protein (MBP). Clinical manifestations were mild intellectual deficit, involuntary movements and ataxia. MRI of the brain showed diffusely abnormal white matter. Visual evoked responses were normal. Central conduction was prolonged on median somatosensory evoked potentials and no central response was seen with posterior tibial somatosensory potentials. Putative congenital deficiency of MBP does not necessarily cause abnormal visual evoked responses.


Stimulation of 5-HT1 receptors in the spinal cord changes substance P-induced behaviour. This study examined whether intrathecal (i.th.) injection of different 5-hydroxytryptamine (5-HT) receptor agonists modulated the behavioural response to substance P. Given intrathecally, substance P produces a behavioural syndrome consisting of biting of the lower parts of the abdomen and reciprocal hindlimb scratching, which may be indicative of nociceptive stimulation. The number of substance P-induced bites was reduced when counted 5 min after intrathecal injection of 5-HT, p-chloroamphetamine (PCA) which causes release of 5-HT from neuronal terminals, the non-selective 5-HT receptor agonist quipazine, the selective 5-HT1 receptor agonists (+)-8-hydroxy-2-(di-n-propylamino)tetralin [(+)-8-OH-DPAT], 5-methoxy-3(1,2,3,6-tetrahydropyridin-4-yl)-1H-indole (RU 24969) and 1(m-chlorophenyl)piperazine (mCPP), but was unchanged by treatment with the 5-HT2/5-HT1C receptor agonist (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI). The number of scratches was significantly increased 5 min after injection of 5-HT and RU 24969. The results showed that intrathecal injection of 5-HT agonists, with a high affinity for the 5-HT1 receptor subtypes, reduced the total number of responses induced by intrathecal injection of substance P, whereas a 5-HT2/5-HT1C receptor agonist did not affect the behavioural response to the intrathecal injection of substance P.


Spinal and intestinal levels of substance P, calcitonin gene-related peptide and vasoactive intestinal polypeptide following perendoscopic injection of formalin in rat colonic wall. Acute inflammation of the colonic wall was induced by perendoscopic injection of formalin in rats. As compared to control animals (no endoscopy, no injection), the procedure was followed by a marked reduction of immunoreactive substance P, calcitonin gene-related peptide and vasoactive intestinal peptide concentrations in rectosigmoid wall. Tissue substance P concentration in the spinal cord, at the level of afferent projection, increased at the same time. The three peptides tested are thus likely to be involved in the pathophysiology of acute intestinal inflammation. In addition, substance P may play a role in the transmission of nociceptive signals from the inflamed colonic segment.


The effect of p-chloroamphetamine and p-chlorophenylalanine on the level of thyrotropin-releasing hormone and its receptors in some brain structures and lumbar spinal cord of the rat. The concentration of thyrotropin-releasing hormone (TRH) and the density and affinity of TRH receptors were examined in the ventral and dorsal lumbar spinal cord, nucleus accumbens and striatum of rats with the 5-hydroxytryptamine (5-HT) nerve terminal destroyed with p-chloroamphetamine (PCA), or in animals treated with the inhibitor of 5-HT synthesis p-chlorophenylalanine (PCPA). PCA (2 x 10 mg/kg i.p., 9 and 8 d before killing) and PCPA (3 x 300 mg/kg i.p., 72, 48 and 24 h before killing)--either of them dramatically diminishing the 5-HT and 5-HIAA concentrations in all the examined structures--reduced the TRH level and increased the density of TRH receptors in the ventral lumbar spinal cord. PCPA also reduced the TRH content in the nucleus accumbens. The PCA-induced reduction in the TRH level and increase in the density of TRH receptors in the ventral lumbar spinal cord were significantly attenuated by citalopram (2 x 20 mg/kg i.p., 30 min before PCA), a selective inhibitor of 5-HT uptake. Our results constitute a further proof that coexistence of TRH and 5-HT takes place in the ventral lumbar spinal cord and then indicate that other form(s) of relationship between 5-HT and TRH may exist in some parts of the central nervous system. They also suggest that an up-regulation of TRH receptors occurs in the spinal cord as a result of TRH depletion.


Primary choroid plexus papilloma located in the suprasellar region: case report. A 34-year-old woman with primary choroid plexus papilloma occurring in the suprasellar region is reported. No connection with the ventricular system was found during intraoperative observations. The findings of pathological examinations such as hematoxylin and eosin staining, transthyretin (prealbumin) immunoreactivity, and electron microscopy were consistent with choroid plexus papilloma. Radiologically, it was extremely difficult to differentiate from tuberculum sellae meningioma. To our knowledge, this is the first case of primary choroid plexus papilloma in this location reported in the literature.


Dose changes in long- and medium-term intrathecal morphine therapy of cancer pain. Intrathecal morphine analgesia for the treatment of cancer pain was administered using implanted ports and drug delivery systems (DDS) in 79 patients. Effective control of the pain was achieved in nearly all patients; in only two patients was the use of the DDS discarded because of relative ineffectiveness. Fifty-three manual drug release systems (41 lumbar, 12 ventricular) and 26 lumbar ports were used. Forty patients survived more than 2 months; the maximum survival time was 560 days (mean survival time, 80 days with a port system, 100 days with a manual DDS). Patients still alive at the time of this study, i.e., with unknown survival time, were excluded. The initial mean daily dose was 8.5 mg in lumbar ports, 2.75 mg in lumbar DDS, and 0.2 mg with intraventricular application. Dose change patterns disclosed no alteration of the initial dose in 18 of 26 port patients, an initial increase in 4, a preterminal increase in 3, and a single intermittent increase in 1 patient. Of 40 lumbar DDS patients, 13 showed a constant dose, 9 an initial, 3 a preterminal, and 5 an intermittent increase. Three patients with less than 2 months' survival time had a rather continuous increase. All long-time survivors (i.e., with more than 2 months' survival time) reached a plateau and remained there until a preterminal if any increase occurred. These findings suggest the morphine dosage to be indicative of the progress of the disease rather than of a drug tolerance.(ABSTRACT TRUNCATED AT 250 WORDS)


Morphological differences between projection neurons of the core and shell in the nucleus accumbens of the rat. The somatodendritic morphology of projection neurons in the shell and core of the rat nucleus accumbens was studied. These cells were retrogradely labelled with Fast Blue from the ventral mesencephalon (substantia nigra/ventral tegmental area) and subsequently injected intracellularly with Lucifer Yellow and processed immunocytochemically. Digitized reconstructions revealed that the cell bodies of neurons located throughout the nucleus are small-to-medium in size. Neurons in the shell have significantly fewer dendritic arbours with fewer branch segments, fewer terminal segments, and lower spine densities than those in the core. Values for the same parameters are significantly greater for cells in lateral than in medial parts of the shell but the same for neurons located within and without enkephalin enriched parts of the core, with an exception of spine density being significantly greater in the enkephalin-rich compartment. Calculations based on these data reveal that neurons in the core have as much as 50% more surface area than those in the shell, which suggests that core neurons have a greater potential for collecting synaptic information than have shell cells. Furthermore, the differential distribution and action of various neurochemicals such as dopamine in the shell and core, supports the idea that different morphologies reflect the presence of distinct neuronal circuits in these two territories.


Alteration in the levels of thyrotropin releasing hormone, substance P and enkephalins in the spinal cord, brainstem, hypothalamus and midbrain of the Wobbler mouse at different stages of the motoneuron disease. The present study was undertaken to quantify selected neuropeptides (thyrotropin releasing hormone, substance P, methionine and leucine enkephalin) in the cervical spinal cord and other regions of the central nervous system of Wobbler mice by radioimmunoassays during several stages of the motoneuron disease compared with age- and sex-matched normal phenotype littermates. In Wobbler spinal cord, thyrotropin releasing hormone is higher early in the disease, whereas in the brainstem it is higher at a later stage. Substance P in spinal cord is also higher late in the disease. Leucine enkephalin levels are greater at all stages in diseased spinal cord and brainstem, but methionine enkephalin increases only late in the disease. Highly significant increases of the peptides (except thyrotropin releasing hormone) appear in hypothalamus and midbrain only late in the motoneuron disease. Regression analyses show that thyrotropin releasing hormone in spinal cord and brainstem decreases normally with age in the control mice and at a faster rate related to the extent of motor impairment in Wobbler mice. Thyrotropin releasing hormone and methionine enkephalin in the Wobbler brainstem correlate (P less than 0.05) with the progress of the motoneuron disease. Methionine enkephalin increases faster in Wobbler brainstem and decreases faster in control spinal cord with age. The increase of leucine enkephalin in the Wobbler spinal cord correlates significantly with age and with the progress of the disease, but leucine enkephalin declines slightly with age in the controls. The changes of substance P in spinal cord and brainstem do not correlate significantly with the progress of the disease. In the hypothalamus, increasing values for substance P in control specimens and enkephalins in Wobbler specimens are significantly correlated with age. However, in the midbrain, higher methionine and leucine enkephalin levels are significantly associated with age only in the control mice. Alterations of neuropeptides in the Wobbler mouse spinal cord and brainstem may result from the degeneration of bulbospinal raphe neurons projecting to the ventral spinal cord, or from primary afferent or interneuronal nerve terminals. The data imply that the neuronal degeneration process in the Wobbler motoneuron disease is not limited to motoneurons. In the spinal cord, the data support our previous hypothesis that neuronal sprouting presynaptic to the motoneurons may account for increased neuropeptide concentrations. Alternatively, synthesis and/or degradation of these peptides may be altered. In addition, it is proposed that enkephalinergic neurons may develop abnormally in Wobbler mice. The early increase of leucine enkephalin in the Wobbler spinal cord possibly indicates its importance in the etiology of the motoneuron disease.


Monoclonal antibody BM89 recognizes a novel cell surface glycoprotein of the L2/HNK-1 family in the developing mammalian nervous system. A monoclonal antibody, BM89, obtained with Triton X-114-treated pig synaptic membranes as an immunogen, recognizes a neuronal antigen in the newborn porcine nervous system. By immunohistochemistry, BM89 staining was observed within the neuropil of all areas of the forebrain and spinal cord tested. In addition, BM89 labeled the cell bodies and proximal dendrites of spinal cord neurons. In the peripheral nervous system, BM89 immunoreactivity was present in a subpopulation of dorsal root ganglion neurons and was predominantly associated with non-myelinated axons in peripheral nerves. Initial biochemical characterization of the antigen in pig brain showed that it is an integral membrane glycoprotein with a molecular weight of 41,000. Moreover, it cross-reacts with the L2/HNK-1 carbohydrate epitope expressed by members of a large family of glycoproteins. Homologous antigens with molecular weights of 41,000-43,000 were identified in the rat, rabbit and fetal human brain. Immunoblotting and immunohistochemistry revealed that the epitope recognized by BM89 is developmentally regulated in the rat nervous system. In cryostat sections from rat cerebellum, spinal cord and dorsal root ganglia, an age-dependent decline of BM89 immunoreactivity was observed during postnatal development. In the cerebellum, the BM89 epitope was very abundant in cells of the external and the internal granular layers between postnatal days 5 and 15. During this period some staining was also identified in the developing molecular layer and the prospective white matter. Subsequently, and in the adult, overall staining was greatly reduced and remaining immunoreactivity was associated only with the internal granular layer. In the spinal cord and dorsal root ganglia, staining was very prominent at postnatal day 5; it decreased considerably thereafter and was barely detectable in the adult. Immunostaining of rat brain and dorsal root ganglion cultures revealed that the BM89 antigen is a cell surface molecule expressed by a subpopulation of central and peripheral nervous system neurons. The biochemical properties in conjunction with the topographical location and the developmental profile of the antigen recognized by BM89 suggest that it may represent a developmentally important recognition molecule.


Amyloidoma of the CNS. II. Immunohistochemical and biochemical study. We present the immunohistochemical and biochemical identification of an amyloidoma localized to the cerebral white matter in a patient who shows no evidence of systemic or extracranial localized amyloid deposits. Immunohistologic and immunochemical studies, using antibodies against biochemically different amyloid fibrils and amyloid-associated proteins, showed reactivity with antibodies only to lambda light chain and serum amyloid P-component. Amino acid sequence analysis of the purified amyloid fibrils extracted from the brain tumor indicates that the amyloid protein is an unusual immunoglobulin lambda light chain, starting at residue five of the variable domain. These fibrils consist of lambda chain fragments of different lengths (10 to 30 kd) very likely arising by polymerization of the amyloid subunit or sequential proteolytic cleavage of the light chain, or both. After exposure to denaturing agents, the 10-kd subunit retains the characteristics of native amyloid fibrils by electron microscopy.


[Arrhythmogenic Takayasu disease].

[Arrhythmogenic Takayasu disease]. The Authors report a case of Takayasu disease in a woman who died at the age of forty-six, in whom the histological examination of the cardio-vascular system revealed giant cells granulomatous arteritis localized in the aortic arch and collateral arteries; endocarditis and granulomatous coronaritis. The bases of arrhythmogenic alterations, in this study, take into account the thrombosis of the conduction system arteriolar vessels and the phlogosis extending to the cardiac plexus.


Selective participation of brain-specific nonhistone Np-3.5 proteins of chromatin in the processes of the reproduction of a defensive habit in response to food in edible snails. The role of brain-specific nonhistone Np-3.5 proteins of chromatin in the processes of the reproduction of a developed defense habit in response to food was studied in preliminarily trained edible snails. It was found that gamma globulins to Np-3.5 over the course of tens of minutes suppress behavioral and neuronal reactions elicited by a specific conditional stimulus, carrot juice, while not altering the reaction to a different conditional stimulus, apple juice. The gamma globulins to other nonhistone proteins of chromatin did not exert an influence on the reproduction of habits of food rejection. It is hypothesized that brain-specific nonhistone Np-3.5 proteins of chromatin are selectively involved in the molecular processes supporting the neurophysiological mechanisms of the extraction of information from long-term memory.


Prevalence of non-A non-B hepatitis and anti-HCV antibodies in a Portuguese dialysis population. Non-A Non-B hepatitis (NANBH) is nowadays one of the most common causes of hepatic dysfunction in dialysis patients. We reviewed the records of 231 HBsAg-negative patients in our unit and found 119 patients with biochemical criteria of NANBH (51.5%), 88 of them (68.9%) with circulating antibodies against HCV (chi 2 P less than 0.0001). Such high prevalence of NANBH was due to an outbreak of NANBH in the late 70s and early 80s. Time on haemodialysis (HD) was the major risk factor of NANBH in this population, and no other risk factors were identified. Prevalence of anti-HCV was similar to reports from non-uraemic populations. Anti-HCV seems to be a reliable test to confirm NANBH, but not better than using common biochemical criteria of NANBH to manage these patients in dialysis units.


NMDA-induced release of nitric oxide potentiates aspartate overflow from cerebellar slices. We report that stimulation of neonatal rat cerebellar slices with N-methyl-D-aspartate (NMDA) release nitric oxide (NO) and also increased the release of aspartate. Inhibition of NMDA receptors with the specific antagonist, 3-((RS)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP) prevented the NMDA-induced release of both NO and aspartate. Similar results were obtained with the inhibitor of NO synthase, NG-nitroargine (NG-ARG). The NO scavenger, haemoglobin prevented the release of aspartate. Under calcium-free conditions NMDA-induced aspartate release was abolished and NO release significantly reduced. These results indicate that NO has a physiological role in the release of aspartate.


Pre-spinal convergence between thoracic and visceral nerves of the rat. Dichotomizing sensory axons have been demonstrated in a number of species and are of significance in understanding the possible mechanisms underlying referred pain. The present study reviews work employing fluorescent dyes as tracers to demonstrate afferent dichotomization in the peripheral nervous system. Dichotomization between the intercostal and splanchnic nerves of the rat was demonstrated by means of intraneural transport of Diamidino yellow or Fast blue. Frequency of pre-spinal somato-visceral convergence averaged 2% (range 0.1-21%). Average frequency of convergence was 8.3% (range 2-23.1%) between internal and external intercostal nerves. Control experiments in which axoplasmic transport was inhibited by vinblastine ruled out the possibility of errors from non-axoplasmic transport of the markers. Thoraco-visceral pre-spinal convergence occurs in the rat and is variable in extent.


The origin of trochlear motoneurons in the larval sea lamprey, Petromyzon marinus L. An HRP study. The origin of trochlear motoneurons in larval lampreys was studied by injection of HRP into the orbit. Immature motoneurons had ventricular attachments, the position of which with respect to the ventricular sulci was used to study their regions of origin. Motoneurons originate from the region between the sulcus intermedius dorsalis and the sulcus intermedius ventralis, which in other parts of the brain were identified as the visceromotor and viscerosensory columns. Both ipsilateral and contralateral immature motoneurons were found. The significance of these findings is discussed in relation to the origins of the trochlear nerve and its nucleus in vertebrates.


Dimethyl sulfoxide (DMSO) blocks GABA-induced current in rat dorsal root ganglion neurons. The effects of dimethyl sulfoxide (DMSO) on gamma-aminobutyric acid (GABA)-induced Cl- currents of rat dorsal root ganglion neurons in primary culture were studied by the whole-cell patch-clamp technique. Bath application of 5 microM GABA evoked sustained inward and outward currents at membrane potentials of -60 mV and +30 mV, respectively, when the external and internal solutions both contained 142 mM Cl-. DMSO at concentrations of 0.3-3% (v/v) caused a dose-dependent inhibition of the inward current induced by 5 microM GABA at -60 mV. DMSO at 3% inhibited the GABA-induced inward and outward currents at -60 mV and +30 mV to similar extents (54% and 53% of the control, respectively), but the time-course of inhibition of the outward current at +30 mV was much faster than that of the inward current at -60 mV. These results suggest that DMSO suppressed the currents by interacting with GABA receptor-Cl- channel complex protein rather than by affecting the lipid-bilayer of the cell membrane.


Non-uniform distribution of GABA activated chloride channels in cultured cortical neurons. Several different neurotransmitter receptors and ion channels exist in non-random distributions in the membrane of nerve and muscle cells, a pattern which appears to have functional implications. The data presented here, based on the distribution of single channels in outside-out patches, directly demonstrate that receptors for gamma-aminobutyric acid (GABA) and their associated chloride channels (GABAA receptors) exist almost exclusively in clusters in the membranes of cultured rat cortical neurons. This non-uniform distribution is present both in innervated and uninnervated neurons.


Identified septohippocampal neurons survive axotomy: a fine-structural analysis in the rat. Previous studies have indicated that interruption of the connections between the medial septum and hippocampus by cutting the axons results in degeneration and death of the projecting septal neurons. However, in these studies cell death has been inferred primarily from the loss of immunoreactivity for transmitter-specific enzymes. In the present study, we labeled septohippocampal projection neurons by retrograde tracing and then cut their axons. Subsequent intracellular injection of prelabeled cells revealed the morphology of the soma and dendrites and allowed us to examine the ultrastructure of these neurons. A large number of septohippocampal neurons survived even 10 weeks after axotomy, suggesting that axotomized septohippocampal neurons survive for considerable periods beyond the time at which they stop expressing transmitter-specific immunoreactivity. Survival of axotomized neurons is a prerequisite for pharmacological interference aimed at reactivating transmitter expression, axonal re-growth, and the eventual reintegration into functionally relevant circuitries.


Development of cutaneous and proprioceptive afferent projections in the chick spinal cord. Muscle and cutaneous nerves were individually labeled with DiI in chick embryos to examine the development of sensory afferent arborizations in the spinal cord. Initially, cutaneous and muscle arbors were similar; both types first entered the spinal gray matter at stage 28-29 (embryonic day (E) 6). Differences in projections were first observed by late stage 34 (E8.5): muscle afferent collaterals extended almost unbranched to the level of motoneuronal dendrites while cutaneous afferents branched frequently and remained within the dorsal horn. Projections of putative small caliber axons into laminae 1 and 2, located laterally in the chick, did not develop until E13-14.


The cuneate nucleus in the rat does have an anatomically distinct middle region. Recently obtained anatomical evidence supports the division of the rat cuneate nucleus (CN) into three rostrocaudal regions, with the middle region receiving a disproportionately greater share of the primary sensory input. The CN in the rat conforms to the basic rostrocaudal CN pattern described in other mammals, including cat, monkey and raccoon.


Effect of clonidine on the release of serotonin from the rat hippocampus as measured by microdialysis. The purpose of the present study is to clarify the effect of clonidine on the release of serotonin from the rat hippocampus in vivo. For this purpose, endogenous serotonin release was measured by brain microdialysis. Potassium-evoked serotonin release from the hippocampus of freely moving rats was significantly inhibited when clonidine (10(-5) M) was added to the perfusion solution, while the 5-hydroxyindoleacetic acid output remained unchanged. In catecholaminergically denervated rats, clonidine (10(-5) M) also inhibited the potassium-evoked serotonin release from the hippocampus and the 5-hydroxyindoleacetic acid output was unaffected by clonidine. These results suggest that the inhibitory effect of clonidine on serotonin release from the hippocampus might reflect the activation of alpha 2-adrenoceptors which are localized on the serotonergic nerve terminals.


Topographic projections from the subiculum to the limbic regions of the medial frontal cortex in the cat. When WGA-HRP was injected into the subicular cortex of the ventral hippocampal formation (Hv) in the cat, terminal labeling was seen ipsilaterally in the caudoventral parts of the medial frontal cortex: in the infralimbic cortex (area 25), ventral part of the prelimbic cortex (area 32), and caudoventral part of the orbitofrontal cortex (area 12). The terminal labeling was observed in all cortical layers except layer 1. The cells of origin of these projections were then confirmed to be pyramidal neurons in the subiculum of the Hv. The projections were topographically organized: the temporal-septal shift along the long axis of the subiculum of the Hv corresponded to a rostroventral-caudodorsal shift in the anterogradely labeled limbic regions of the medial frontal cortex.


Serotonin uptake inhibition: in vivo effect of sertraline in rats. Sertraline, a potent and selective serotonin uptake inhibitor, was used to analyze the changes occurring in the serotonin system after uptake inhibition in vivo. Sertraline (11 mg/kg) lowered extracellular 5-hydroxyindolacetic acid (5-HIAA), measured in rat hippocampus by in vivo voltammetry, for about 3 h. The interaction between sertraline and drugs known to interfere with the release or uptake of serotonin (L-5-hydroxytryptophan (5-HTP), d-norfenfluramine and tianeptine) was then studied. The sertraline-induced decrease in extracellular 5-HIAA was related to the inhibition of uptake.


Experimental peripheral neuropathy decreases the dose of substance P required to increase excitatory amino acid release in the CSF of the rat spinal cord. Partial ligation of the sciatic nerve of rats produces hyperalgesia similar to that seen in humans following nerve injury. In this study, we used microdialysis of the spinal cord cerebral spinal fluid (CSF) to test the hypothesis that hyperalgesia is due to an enhanced release of excitatory amino acids (EAA) in response to substance P (SP). Intrathecal SP caused release of aspartate and glutamate in the CSF of rats with partial sciatic ligation at a dose of SP that did not cause release in sham operated animals. Neonatal capsaicin pretreatment blocked SP-induced EAA release in both sham and sciatic ligated animals. Release of EAAs in ligated animals was not significantly different from release in sham-operated animals following higher doses of SP or chemical nociceptive stimulation. These results demonstrate a partial sciatic ligation-induced decrease in the dose of SP required to initiate EAA release in the CSF of the spinal cord, a change which could play an important role in hyperalgesia.


Vocalization and marked pressor effect evoked from the region of the nucleus retroambigualis in the caudal ventrolateral medulla of the cat. It is well established that the nucleus retroambigualis (NRA) of the cat contains a population of expiratory-related neurons. We report here that in the unanesthetized, decerebrate cat, microinjections of 300-900 pmol of D,L-homocysteic acid within the NRA evoked excitation of laryngeal as well as expiratory muscles, and often pressor responses. Moreover, vocalizations, which did not sound like normal feline vocalizations (i.e., hiss, howl, mew, growl), were evoked from a restricted region of the NRA, 1-3 mm caudal to the obex. The results indicate that in addition to its role in expiration: (i) the NRA plays an important role in the control of laryngeal muscles and the production of vocalization; and (ii) that neurons in the NRA region can modulate arterial pressure.


Regional cerebral protein synthesis after transient ischemia in the rat: effect of the AMPA antagonist NBQX. Normothermic rats with 12 min, complete cerebral ischemia were treated with the AMPA antagonist 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo (F) quinoxalinedione (NBQX) [10], which prevents CA1 pyramidal neuron loss. Twenty hours after ischemia, cerebral protein synthesis rate (CPSR) was measured autoradiographically using [35S]methionine. Ischemia caused a 38% decrease of CPSR in CA1, and postischemic treatment with NBQX caused a 66% decrease in this region. Also treatment with NBQX alone resulted in a decrease (22% in CA1) of the CPSR. Since some evidence exists that the neuroprotective effect of NBQX is related to blockade of the fast AMPA-mediated transmission, the further decrease of the postischemic CPSR in CA1 could be a mere side effect.


Autoradiographic localization of [125I]charybdotoxin binding sites in rat brain. Charybdotoxin, a 37 amino acid peptide isolated from scorpion venom, is a potent inhibitor of potassium channel function. [125I]charybdotoxin was originally believed to be a selective ligand for the Ca(2+)-sensitive channel in many tissues, but it appears to bind only to a voltage-sensitive potassium channel in brain. We found high densities of [125I]charybdotoxin binding in lateral olfactory tract, interpeduncular nucleus and a variety of mesencephalic nuclei. Moderate levels were found in the cerebral cortex, medial thalamus, hypothalamus and selected thalamic nuclei. These results indicate that [125I]charybdotoxin identifies a potassium channel or channels with a unique distribution in the brain.


Galanin and bethanechol appear to activate the same inwardly rectifying potassium current in mudpuppy parasympathetic neurons. Galanin- and bethanechol-activated whole cell currents were studied in dissociated mudpuppy parasympathetic neurons kept in 12.5 mM KCl. The bethanechol-induced current was reduced when generated during a galanin-induced current. The galanin and bethanechol currents reversed at -45 mV and exhibited inward rectification at more negative potentials. Both the galanin- and bethanechol-induced current relaxations recorded during negative voltage steps were fitted best by two exponentials with the averaged time constant values not significantly different for the galanin and bethanechol currents. We conclude that galanin and bethanechol activate a similar membrane potassium conductance which is different from the background potassium conductance.


The effect of chronic clozapine on basal dopamine release and apomorphine-induced DA release in the striatum and nucleus accumbens as measured by in vivo brain microdialysis. Previous studies have produced conflicting results concerning the effect of chronic oral vs. parenteral (i.p.) clozapine administration on dopamine (DA) release and metabolism in the striatum and nucleus accumbens (n. accumbens) of freely moving rats using in vivo microdialysis. In this study, parenteral chronic clozapine (20 mg/kg/day for 21 days, i.p.) had no effect on basal DA release and metabolism in either region. Chronic treatment with parenteral clozapine also did not reverse the decrease in DA release and metabolism in striatum and n. accumbens produced by apomorphine (100 micrograms/kg, s.c.). These results differ significantly from a previous report following i.p. clozapine and confirm the results previously reported with oral clozapine.


Chemical identification and morphological characterization of the inferior olive in the frog. Tritiated D-aspartate was injected into the cerebellar cortex of grassfrogs, Rana temporaria. Retrograde labeling was observed in a cell column of the contralateral caudal medulla, but not in other areas known to give rise to cerebellar mossy fibers. The aspartate-positive neurons are therefore considered to represent the origin of cerebellar climbing fibers in the inferior olive, as reported earlier for rat and turtle by other investigators. Contrary to earlier reports we found no extraolivary climbing fibers in the VIIIth nerve and in Scarpa's ganglion. Our results support the view that the climbing fiber system of vertebrates is anatomically, physiologically and chemically very distinct and phylogenetically very conservative.


The GABAA receptor complex in experimental absence seizures in rat: an autoradiographic study. The regional distribution of radioactive ligand binding for different receptors of the gamma-aminobutyric acid A (GABAA)-benzodiazepine-picrotoxin chloride channel complex was measured on tissue section by autoradiography in brains taken from a genetic strain of Wistar rats with spontaneous absence-like seizures, the genetic absence epilepsy rats from Strasbourg (GAERS), and a control colony. The ligands employed included [3H]muscimol for high affinity GABA agonists sites; [3H]SR 95531 for the low-affinity GABA sites; [3H]flunitrazepam for the benzodiazepine sites; and [35S]t-butyl bicyclophosphorothionate (TBPS) for the picrotoxin site. There was no significant change between GAERS and control animals in [3H]flunitrazepam and [35S]TBPS binding. However, there was significantly decreased [3H]muscimol and [3H]SR 95531 binding in the CA2 region of the hippocampus of the GAERS. This was due to a decrease in Bmax of both [3H]muscimol and [3H]SR 95531 binding in the epileptic strain.


HT7, Neurothelin, Basigin, gp42 and OX-47--many names for one developmentally regulated immuno-globulin-like surface glycoprotein on blood-brain barrier endothelium, epithelial tissue barriers and neurons. The antigens HT7 and Neurothelin were recently described as inducible membrane glycoproteins on chick blood-brain barrier endothelium. Our results demonstrate that the Neurothelin antibody 1W5 does recognize the isolated HT7 protein as well as the HT7 protein expressed by COS-1 cells transfected with the HT7 cDNA. Therefore, the HT7 and Neurothelin epitopes can be found on an unique glycoprotein. By low stringency hybridization we have isolated the murine cDNA clone encoding the homologous murine protein. The nucleotide sequence of the open reading frame is identical to that of the cDNA clones encoding the gp42 protein and the Basigin antigen expressed in F9 cells and in the early mouse embryo. The murine amino acid sequence shows 94% identity with the rat OX-47 membrane glycoprotein which was shown to be expressed in blood-brain barrier endothelium and several epithelial cell layers. These results demonstrate that HT7, Neurothelin, gp42, Basigin and OX-47 are different names for probably the same glycoprotein in different species.


Changes in striatal dopamine release and metabolism during and after subchronic haloperidol administration in rats. The release and metabolism of dopamine (DA) in the striatum of rats during and after subchronic haloperidol (HAL) administration (3 weeks) was assessed using in vivo microdialysis. Basal extracellular levels of DA, DA metabolites (homovanillic acid and 3,4-dihydroxyphenylacetic acid) and the serotonin metabolite (5-hydroxyindoleacetic acid) were not significantly different from control values at 3 weeks of HAL administration and 3 days after drug withdrawal. The specific DA D2 receptor antagonist, raclopride (0.5 mg/kg, i.p.), significantly increased DA release and metabolism in control animals, but decreased DA release in the HAL-treated groups at 3 weeks of drug treatment. This effect was not significant following drug withdrawal. These results contrast with our previous finding that chronic HAL treatment (32 weeks) increases basal DA metabolism and further support the possibility that changes in DA function differ following short term vs. long term neuroleptic exposure.


Influence of electro-acupuncture on the release of substance P and the potential evoked by tooth pulp stimulation in the trigeminal nucleus caudalis of the rabbit. The effects of electro-acupuncture (EAP) on the release of substance P (SP) and the responses evoked by tooth pulp stimulation (ST) in superficial layers of the trigeminal nucleus caudalis (Vc-I,II) were studied in rabbits. ST evoked increase in release of immunoreactive SP (iSP). This increase was inhibited by EAP in 9 of 13 animals. The potentials evoked by ST were composed of two main components with latency times of ca 4.3 msec and ca. 9.4 msec. The latter component, reflecting the excitation of A delta fibers, was significantly inhibited by CP-96,345 (3 mg/kg, i.v.), an SP antagonist. EAP also inhibited the latter component in 8 of 11 animals. These results suggest that one of the mechanisms of analgesia induced by EAP is inhibition of stimulus-evoked SP release in the Vc-I,II.


A heparin-binding protein in porcine seminal plasma stimulates neurite outgrowth on neuroblastoma cells in culture. A protein of neurite outgrowth activity has been identified in porcine seminal plasma after ammonium sulfate precipitation and affinity chromatography on heparin-Sepharose. Upon SDS-PAGE, the polypeptide is shown to have a M(r) of 16,000-18,000. Biologically by induction of neuritic processes on neuroblastoma cells, and immunologically by cross-reaction with specific antisera, this seminal plasma protein differs from acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF) and nerve growth factor (NGF). The neurite outgrowth activity is relatively stable at pH 3-7 and under denaturing conditions of 8 M urea and beta-mercaptoethanol, but is inactivated by treatment of trypsin. This appears to be a novel protein, enhancing morphological differentiation of neuroblastoma cells in culture.


Estimating the time of death in stillborn fetuses: I. Histologic evaluation of fetal organs; an autopsy study of 150 stillborns. To determine whether the autopsy histology of fetal tissues can determine the time of death of stillborn fetuses. Hematoxylin and eosin slides from autopsies of 150 stillborn fetuses with well-timed deaths were evaluated retrospectively. Fetuses were divided into a learning set (100 fetuses) and a test set (50 fetuses). From assessment of the 100 fetuses in the learning set, 23 histologic features were identified with possible temporal associations with fetal death. When those histologic features were randomly and blindly assessed in the 50 test fetuses, ten features performed well as diagnostic tests (sensitivity, specificity, and positive predictive values at or above 0.875), correctly classifying 43 of 50 fetuses (86%) with respect to the time of death. The ten histologic features and their predicted death-to-delivery intervals were: loss of nuclear basophilia in individual cells in renal cortical tubules (4 hours), liver (24 hours), inner half of the myocardium (24 hours), outer half of the myocardium (48 hours), bronchial epithelium (96 hours), and tracheal cartilage (1 week); and loss of nuclear basophilia of all cells in the liver (96 hours), gastrointestinal tract (1 week), adrenal (1 week), and kidney (4 weeks). The development of these histologic changes appeared to be accelerated by fetal hydrops and a delivery-to-autopsy interval exceeding 24 hours and decelerated by fetal gestational age under 25 weeks. Histologic changes identifiable in hematoxylin and eosin-stained fetal tissue may be useful for estimating the time of death in many stillborn fetuses.


Placenta accreta/percreta/increta: a cause of elevated maternal serum alpha-fetoprotein. To investigate the relationship between elevated maternal serum alpha-fetoprotein (MSAFP) and abnormal placental adherence (placenta accreta/percreta/increta). We reviewed the MSAFP levels of 11 women who had cesarean hysterectomies because of placenta accreta/percreta/increta. The control group consisted of 14 women who delivered by cesarean because of placenta previa but who had no abnormal placental adherence. Five of the 11 women with placenta accreta/percreta/increta had elevated MSAFP, whereas all 14 controls had normal levels. These results indicate a significant association between elevated MSAFP and placenta accreta/percreta/increta (P = .017). Patients with an unexplained elevation of MSAFP as well as placenta previa may be at increased risk for abnormal placental adherence.


Low birth weight and preterm delivery in relation to early-gestation vaginal bleeding and elevated maternal serum alpha-fetoprotein. To determine whether early-gestation vaginal bleeding and elevated maternal serum alpha-fetoprotein (MSAFP) are independent risk factors for adverse infant outcomes. We conducted a cohort study of 201 women with an elevated MSAFP (at least 2.0 multiples of the median [MOM]) and a second-trimester ultrasound evaluation at Swedish Hospital Medical Center between January 1989 and March 1991, and 211 women with MSAFP levels below 2.0 MOM who had also undergone ultrasound evaluations during the same period. Stratified analyses demonstrated that early-gestation bleeding and elevated MSAFP were independent risk factors for the delivery of both preterm and low birth weight infants. Compared with women with no history of early-gestation bleeding or elevated MSAFP, women with early-gestation bleeding alone had a relative risk (RR) of preterm delivery of 4.3 (95% confidence interval [CI] 1.5-12.1); non-bleeders with elevated MSAFP had an RR of 4.6 (95% CI 1.9-10.9). A combined history of early-gestation bleeding and elevated MSAFP was associated with an almost sixfold increased risk of preterm delivery (RR 5.8; 95% CI 2.2-15.6). Analyses restricted to women with normal-appearing appearing placentas by second-trimester ultrasound evaluations yielded similar results. These findings support an earlier report documenting the independence of early-gestation bleeding and elevated MSAFP as predictors of adverse infant outcomes.


Effect of dextran on predamaged corneal endothelium: an organ culture study. Dextran T500 was used as a supplement in organ culture medium to evaluate its effect on corneal endothelium with disseminated damage. After 1 week of culturing, the control group showed an intact endothelium throughout the entire cornea. However, in the dextran-supplemented group, multiple endothelial necroses were observed in 5 of 10 corneas. Besides, intracytoplasmic vacuoles and granules, deposits on the endothelial surface and widened intercellular spaces were quite numerous. The potential toxicity of dextran to degenerated corneal endothelial cells may be responsible for these findings.


[Comparative study of advanced-stage Hodgkin's lymphoma patients treated with either COPP or ABVD drug combinations]. Between January 1985 and March 1988, 45 patients with advanced Hodgkin's disease were randomly assigned to receive either COPP (22 patients) or ABVD (23 patients). 9 and 16 patients achieved complete remission in the COPP and ABVD groups, respectively. One of the complete responders (COPP) relapsed during the follow-up. Patients treated with COPP experienced myelosuppression and neurotoxicity more frequently whereas alopecia and gastrointestinal symptoms were observed among those receiving ABVD. Present data confirm the favorable impact of primary ABVD treatment in advanced Hodgkin's disease.


[The use of endoprostheses (stents) in airway stenosis]. Six patients with obstruction of the trachea were treated with silicone rubber endothracheal stents implanted with flexible bronchoscope. In every case the stent caused significant clinical improvement of the ventilation. At the postintubation stenosis the stent can result a final recovery, at the malignant processes the implantation seems to be a new palliative method.


[Secondary acute leukemia in Hodgkin's disease]. Two secondary acute leukaemias out of 166 patients with Hodgkin's disease were detected. At a young female patient treated only with chemotherapy the acute leukaemia developed 39 months following the diagnosis of Hodgkin's disease. Phenotype of leukaemic blast cells could not be determined exactly. After a short complete remission (1,5 month) the patient died because of the progression of her leukaemic process. The other young male patient treated with radio- and chemotherapy had six years disease free interval between the diagnosis of two malignant diseases. The combined cytostatic treatment of his acute lymphoblastic leukaemia resulted complete remission that lasted for one year but later the progression of leukaemia caused death. In connection with these cases the authors reviewed the current questions of treatment related acute leukaemias in Hodgkin's disease.


[Implantation of a Stecker metal prosthesis in malignant stenosis of the gastric stump]. A 55 year old patient 3 years after gastric resection (Billroth II) has got gastric outlet stenosis. Half year ago was established a tumour recidive, that the time of reoperation was inoperable. Under endoscopic and radiological control we have introduced and placed a 19 mm diameter, 4 cm long balloon expandable tantalum stent (Strecker stent). The gastric passage has normalised and after 6 month the patient is symptom free.


[Correlation between elevated maternal serum alpha-fetoprotein, certain pregnancy complications and fetal death]. In a prospective epidemiological study of 23,792 singleton pregnancies the authors investigated the relationship between elevated maternal serum alfa-fetoprotein level and the risk of pathological pregnancy outcome. It has been found that elevated level of maternal alfa-fetoprotein (above 2.5 MoM) is associated with a higher risk of pathological pregnancies and an increased risk of subsequent fetal death. Physicians should be aware of the benefits of the maternal alpha-fetoprotein screening not only in the context of prenatal detection of congenital defects, but also in early identification of increased risk of certain obstetric complications.


[Initial results of hybrid chemotherapy of Hodgkin's disease using COPP/VBA]. 19 patients with advanced, previously untreated Hodgkin's disease were treated with COPP/ABV (cyclophosphamide, vincristine, procarbasine, prednisolone, adriamycin, bleomycine, vinblastine) hybrid regimen. Complete response was achieved in 8 patients, partial response in 9 patients and primary treatment failure occurred in 2 patients. Two patients received involved field irradiation for the residual disease. 17 months after finishing the therapy one of the complete responders suffered a relapse. Of the 8 partial responders, 5 patients relapsed. 2 relapses occurred during the treatment and 3 relapses were observed in the first 7 months of follow-up. 2 of the partial responders and both nonresponders succumbed to their disease. There were no serious side effects and treatment-related death did not occur.


Overexpression of the A9 antigen/alpha 6 beta 4 integrin in head and neck cancer. A tumor-associated antigen detected by monoclonal antibody UM-A9 raised against a cultured cell line from a patient with an aggressive SCC of the oral cavity has been defined. The A9 antigen is abnormally expressed in squamous cancers, with loss of basal polarization and increased intensity of expression distinguishing malignant from normal cells. A minority of cultured SCC cell lines and about one third of fresh tumors exhibit polarized A9 expression. The increased intensity and loss of polarized expression of A9 antigen in recurrent and metastatic tumor cell lines when compared with primary or early tumor cell lines from the same patients indicated an association of altered expression with tumor progression. When A9 expression was evaluated in frozen tumor sections, three patterns of expression representing increasing intensity and loss of polarization were observed. Patients whose tumors exhibited the most intense A9 antigen expression had a higher rate of early relapse than patients whose tumors exhibited low intensity and polar expression. Loss of blood group antigen expression was also associated with poor prognosis, and together high A9 antigen expression and loss of blood group defined a group of patients at high risk of early relapse. The A9 antigen is immunologically and biochemically identical to the alpha 6 beta 4 integrin. The association of high expression of the A9/alpha 6 beta 4 integrin as a prognostic factor is supported by similar findings with a mouse model system. The mouse tumor-associated antigen, TSP-180, which is also an alpha 6 beta 4 integrin, distinguishes highly metastatic tumor cells from nonmetastatic variants of the same tumor line. In SCC, the alpha 6 beta 4 integrin contributes to attachment to laminin since anti-alpha 6 subunit specific antibody blocks cell attachment and only the beta 4 subunit is found in association with the alpha 6 subunit in these cells. Similar findings were obtained in colon carcinomas. Antibodies and peptides that block laminin attachment may lead to the development of antimetastatic agents for squamous carcinomas. The beta 4 subunit is unique from other integrins in that it has an unusually long cytoplasmic domain, the function of which is not known. The beta 4 subunit is heavily phosphorylated under conditions that favor anchoring and terminal differentiation in normal keratinocytes. Paradoxically the beta 4 subunit is also heavily phosphorylated in tumor cells, which are highly migratory and immortalized.(ABSTRACT TRUNCATED AT 400 WORDS)


An overview of molecular biology in otolaryngology. The molecular biology revolution is just beginning to bear fruit in medicine. This is an exciting time to be active in the field of biomedical research. It is also an exciting time for clinicians, watching the applications of our new-found knowledge to clinical specialties.


[Investigations of the ototoxicity of dicortinef (ear drops)]. The authors investigated the ototoxic influence of Dicortinef in laboratory animals. Their examinations were performed on 15 guinea pigs (weighing 210-380 g.) after application of this medicine to the fenestra rotunda. The harmful effect of Dicortinef was expressed by the characteristic fall in the microphonic potential (PM) and potential of the acoustic nerve (AP).


Cardiac function in obstructive sleep apnea patients following uvulopalatopharyngoplasty. Obstructive sleep apnea syndrome (OSAS) is associated with severe cardiac arrhythmias and conduction abnormalities. Cor pulmonale and right-sided heart failure may ensue. Uvulopalatopharyngoplasty (UPPP) is one of several treatment modalities suggested for OSAS. Tracheotomy and CPAP treatment in adult OSAS patients and adenotonsillectomy in children with OSAS were shown to lead to improvement in some cardiac parameters. Cardiac function was prospectively evaluated in 19 OSAS patients before and after UPPP. No significant changes after surgery were noted on electrocardiographic studies. Improvement in global and regional function of both ventricles was seen in 91% of the patients. A trend toward significant elevation in left ventricular ejection fraction and a statistically significant increase in right ventricular ejection fraction were observed (45% +/- 9% to 50% +/- 7% [p = 0.007]). Our results support performance of UPPP in selected OSAS patients for relief of potentially life-threatening cardiac pathologies.


The establishment of human T cell lines reactive with specific periodontal bacteria. Human T cell lines (TCLs) were obtained by stimulation of peripheral blood mononuclear cells (PBMCs) with the 2 periodontopathic bacteria, Porphyromonas gingivalis FDC-381 and Fusobacterium nucleatum FDC-263. After the first round of stimulation and rest, the cells responded specifically to the bacteria originally used to establish each line. Throughout the culture period, the responsiveness of each of the TCLs to their specific bacteria increased. Phenotypic analysis of the TCLs revealed heterogeneity of cell types. In both TCLs approximately 80% of the cells were T cells, all of which bore the alpha beta T cell receptor. The P. gingivalis-reactive TCL (PG-TCL) showed approximately equal proportions of CD4+ and CD8+ cells, whereas the F. nucleatum-reactive TCL (FN-TCL) was predominantly CD4+. The expression of CD25, HLA-DR, CD45RA and CD29 on these CD4+ cells varied throughout the culture period of 45 days. These results demonstrate that it is possible to establish long-term T cell lines reacting to specific periodontopathic bacteria.


Hepatitis C core antibody detection in acute hepatitis and cirrhosis patients from Tunisia. The detection of anti-Hepatitis C virus (HCV) Core antibodies is an important addition to HCV antibody testing. In this study it appears to be more specific than the first generation HCV tests and in combination with the detection of anti-C33c antibodies, it is possibly more sensitive. In Tunisia hepatitis C virus is implicated by the presence of anti-Core antibodies in only 8% of the adult cases of acute hepatitis as opposed to 60% for Hepatitis B virus (HBV) and 4% for Hepatitis A virus (HAV). In contrast to the low prevalence of HCV infection among acute hepatitis cases, HCV infection is implicated in 35% of the cirrhosis cases. These results stress the potential importance of HCV infection in the development of cirrhosis which is a relatively common disease in North Africa.


[Cytokeratins, markers of epithelial cell differentiation: expression in normal epithelia]. Intermediate filaments, the most stable of cytoskeleton components, are extremely diverse and usually correlate with the histological subtype since in nearly all cell types a single type of intermediate filament (IF) is found. The cytokeratins, which are specific of epithelia, are the largest and most diverse class of intermediate filaments. Twenty different cytokeratin polypeptides have been identified in humans and separated on the basis of isoelectrical pH and apparent molecular weight using two-dimensional electrophoresis. These data have been used to establish a cytokeratin catalogue which currently serves as a reference [43, 48]. The number of cytokeratin polypeptides expressed ranges from 2 to 5 for each epithelial cell and from 2 to 10 for each epithelium and even of each cell layer within a given epithelium. A broad spectrum of anticytokeratin antibodies with subgroup or single polypeptide specificity is currently available. The distribution of cytokeratins in normal epithelia is reviewed herein and commercially available anti-cytokeratin antibodies are listed.


[Expression of cytokeratins during embryogenesis and in pathologic epithelia]. Epithelial cell intermediate filaments, or cytokeratins, are excellent markers for cell differentiation. During embryogenesis, cytokeratins specific of a stage of differentiation step always become detectable before corresponding morphologic changes: for instance, cytokeratins 5 and 14 are found around the eight week, shortly before stratification of the epithelium occurs, and cytokeratins 1 and 10 are produced before morphologic evidence of keratinization becomes detectable. Among potential diagnostic applications, analysis of cytokeratin patterns of epidermal cells desquamated in the amniotic fluid may provide earlier and less invasive diagnosis than fetoscopic biopsies. Similarly, a review of cytokeratins expressed in a variety of epithelial diseases (involving the epidermis, digestive tract, respiratory tract, urogenital tract, or breast) demonstrated persistence of the original tissue pattern in some instances (this was the case for the majority of simple epithelia) but not in others (complex epithelia). This suggests that cytokeratins may prove valuable as markers for specific tumor stages or types and may provide earlier information than morphologic studies.


Mutations within the S gene of hepatitis B virus transmitted from mothers to babies immunized with hepatitis B immune globulin and vaccine. A variant of hepatitis B virus (HBV) having a specific mutation within the S gene has been found to infect vaccinees. To know whether similar variants were involved in Japan, we analyzed two cases of maternal transmission of HBV in infants immunized with hepatitis B immune globulin and hepatitis B vaccine. DNA clones of HBV S genes were propagated from patients and family members and sequenced. In one family, the DNA clones from the baby patient had a Gly-to-Arg mutation at the 145th codon of the S gene, whereas those from her mother had no such mutations. In the other family, all the DNA clones obtained from the two infected children had the 145th codon intact, but they had a missense mutation at the 126th codon of the S gene, causing an amino acid substitution of Asn for Thr or Ile. This same mutation was observed in 12 of 17 clones of DNA obtained from their mother. In comparison with the wild type HBV-derived hepatitis B surface antigen, the two types of S gene mutations, either at the 145th or the 126th codon, were associated with a significant decrease in the antigenicity of some determinants on the hepatitis B surface antigen, measured by MAb. Amino acid substitution at these sites, therefore, would have induced the escape from conventional vaccines that were S gene products of wild type HBV and also from hepatitis B immune globulin, whose main components were probably also antibodies against the S gene products expressed by wild type HBV.


The in vitro effects of stem cell factor and PIXY321 on myeloid progenitor formation (CFU-GM) from immunomagnetic separated CD34+ cord blood. Two novel cytokines, stem cell factor (SCF) and PIXY321 (a fusion protein, granulocyte macrophage colony-stimulating factor+IL-3), have recently been demonstrated to enhance in vitro adult myelopoiesis. In this study, we compared the success of separating very early hematopoietic progenitor cells (CD34+) from both cord blood (CB) and adult bone marrow (ABM) and their differential response to SCF, PIXY321, and other later-acting colony-stimulating factors (CSF). Briefly, CD34+ cells were isolated from CB and ABM with an anti-CD34 MAb, HPCA-1, and incubated with various combinations of SCF, PIXY321, and other CSF. The percentage of CD34+ cells was decreased in CB compared to ABM before separation (0.54 versus 1.71%) (p = 0.05). Isolated CD34+ cells from CB and ABM were similar in lineage with respect to CD38, HLA-DR, CD33, and CD5, but decreased in CB with respect to B-lineage expression (CD19, CD10, and CD22) (p = 0.05). SCF increased colony forming unit-granulocyte-macrophage (CFU-GM) formation from CB CD34+ cells compared to unconditioned media and had a significant additive increase with IL-3 (p = 0.006) and granulocyte colony-stimulating factor (p = 0.03). SCF also had an additive increase in CB CFU-GM formation with PIXY321 (p = 0.007). PIXY321 had a similar increase in CFU-GM formation from both CB and ABM CD34+ cells compared to the combination granulocyte macrophage colony-stimulating factor + IL-3. When SCF was added to IL-3, PIXY321, or PIXY321 + IL-6, there was an increase in CFU-GM from CB versus ABM CD34+ cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Population-based study of the incidence, complexity, and severity of neurologic disability among survivors weighing 500 through 1250 grams at birth: a comparison of two birth cohorts. Mortality and incidence, complexity, and severity of early childhood neurodevelopmental disability are reported for two cohorts of preterm infants of 500 through 1250 g birth weight. Comparing 1978-1979 (cohort 1) and 1988-1989 (cohort 2), 1-year survival improved from 82 (36%) of 226 to 197 (67%) of 291. Cohort 1 survivors were heavier and more mature than cohort 2 survivors (1047 g vs 930 g, 29.6 vs 27.3 weeks). Parental demographic variables were similar. The incidence of specific disabilities with greater than 97% follow-up to 1.5 years adjusted age did not change: cerebral palsy, 14 (17%) vs 20 (10%); vision loss, 5 (6%) vs 9 (5%); mental retardation, 9 (11%) vs 13 (7%); hearing loss, 3 (4%) vs 7 (4%); and convulsive disorders, 2 (2%) vs 3 (2%). The overall number of disabled children (17 [21%] vs 30 [15%]), complexity of disability (> or = 2 disabilities per child: 11 [13%] vs 10 [5%]), and severity of disability (projected dependency: 6 [7%] vs 10 [5%]) did not differ between cohorts 1 and 2. The cerebral palsy prevalence, based on neonatal survival, dropped from 157 per 1000 to 93 per 1000. Analysis by birth weight-specific categories in 250-g increments did not alter results, but disability rates were highest for those of lowest weight. In contrast to other reports this population-based North American study from a well-developed perinatal regional program reports no increase in incidence, complexity, or severity of disability in preterm infants weighing 500 through 1250 g at birth.


cAMP and Ca2+ act co-operatively on the Cl- conductance of HT29 cells. Previous studies in HT29 cells have revealed that the Cl- channels induced by cAMP or by increasing cytosolic Ca2+, e.g. by addition of ATP, and by hypotonic cell swelling share in common all examined properties, such as ion selectivity and blocker sensitivity. In addition, it was shown that conductances induced by either pathway were not additive. Therefore all three pathways apparently act on the same type of small conductance Cl- channel. In CFPAC-1 cells the general properties of the Cl- conductance were identical. However, the cAMP response was absent. In both cell types the Ca(2+)-mediated conductance response was transient. Here we examine the kinetics of the conductance increases induced by neurotensin (NT, 10(-8) mol/l) or ATP (10(-5) mol/l) in HT29 and CFPAC-1 cells using the slow (nystatin) or fast whole cell patch clamp technique, and we ask whether cAMP influences these kinetics. In the continuous presence of NT the conductance response in both cell types was very transient. It collapsed with a time constant (tau) of 39 (30-56 s) in HT29 and of 33 (27-41 s) in CFPAC-1 cells. The ATP response was also transient with a tau of 49 (42-57 s) in HT29 cells and 102 (77-152 s) in CFPAC-1 cells. Pre-treatment by membrane permeable cAMP (10(-3) mol/l) enhanced the baseline conductance in HT29 but not in CFPAC-1 cells. Furthermore, the ATP- and NT-induced conductance increases became significantly less transient in HT29 but not in CFPAC-1 cells.(ABSTRACT TRUNCATED AT 250 WORDS)


[Pregnancy in insulin-dependent diabetic patients]. There has been a dramatic reduction in perinatal mortality and the incidence of malformation among the offspring of mothers with IDDM, current perinatal infant mortality now being no worse among mothers treated at the best Scandinavian diabetes centres than among non-diabetic mothers (Table 1). Such improvement is dependent upon very well organised care facilities and information services, close cooperation with the diabetic gravida, and the availability of good medical treatment that does not unduly burden her economy. Among diabetic women, careful planning for pregnancy improves the chances of giving birth to healthy offspring.


Competitive binding of a monoclonal antibody SZ-21 with anti-PLA1 antibodies and its potential for clinical application. Both intact IgG and Fab fragments of a monoclonal antibody SZ-21, directed against platelet glycoprotein (GP) IIIa, inhibited binding of anti-PLA1 antibodies to PLA1-positive platelets in a dose dependent manner. Conversely, the binding capacity of the platelets for SZ-21 was decreased in the presence of anti-PLA1 antibodies. In Western blots, the determinant for SZ-21 was present on GPIIIa from either PLA1 or PLA2 homozygotes, but SZ-21 did not affect the interaction of anti-Yukb alloantibodies with their target antigen on GPIIIa. These results suggest that SZ-21 reacts with an epitope on the GPIIIa molecule very close to but not identical with that for anti-PLA1. The existence of PLA1-reactive alloantibodies in serum could be demonstrated by their competitive effect on the binding of SZ-21.


Hodgkin's disease, clinical stages I, II A-B and IIIA. Results of brief chemotherapy followed by irradiation. From October 1980 to September 1985, 152 patients with Hodgkin's disease (HD) in clinical stages (CS) I, II A-B and IIIA were treated with combined modality therapy (CMT): brief chemotherapy (CT) followed by radiotherapy (RT). CS IA and IIA cases received 3 cycles of MOP, while CS IB, IIB and IIIA cases were randomly assigned to receive MOP or MOP alternating with ABVD (4 cycles). Irradiation was delivered according to the areas initially involved and response to CT. CS I and II subjects in complete remission (CR) received localized fields, whereas CS I, II and IIIA subjects in partial remission (PR) received extended fields, with subtotal nodal RT for CS I and II and CS IIIA without pelvic involvement and total nodal RT for CS I and II below diaphragm and CS IIIA with pelvic involvement. Following CT, CR was 60% (IA-IIA: 71.5%, IB-IIB: 37%, IIIA: 55.5%), PR 35.5% and failure 4.5%. After CT and RT, CR was 98% (IA-IIA: 100%, IB-IIB: 95.5%, IIIA: 94.5%). Responses were similar using MOP and MOP/ABVD regimens. Fifteen patients relapsed (10%) and 15 died, 11 due to HD and 4 due to other causes, while after 7 years overall survival and relapse free survival were respectively 87% and 82% (IA-IIA 90% and 85%, IB-IIB: 80% and 80%, IIIA: 87% and 62%). Results were equivalent irrespective of CT regimen.(ABSTRACT TRUNCATED AT 250 WORDS)


Structure of the human DNA repair gene HAP1 and its localisation to chromosome 14q 11.2-12. Apurinic/apyrimidinic (AP) sites are pre-mutagenic DNA lesions which occur spontaneously and following exposure of cells to ionising radiation or chemical mutagens. HAP1 (Human AP endonuclease 1), the major enzyme in human cells initiating repair of AP sites, shows strong sequence homology to DNA repair enzymes from bacteria, Drosophila and other mammalian species. We have cloned the HAP1 gene and determined its complete nucleotide sequence. The site of transcription initiation has been mapped to 452 bp upstream of the ATG initiation codon in the genomic DNA. The HAP1 gene consists of five exons and is unusually small (less than 2.6 kb from transcription initiation site to polyadenylation sequence) with 54% of the protein coding region and the entire 3' untranslated region contained within a single exon. The first exon is non-coding. Regions of three exons show sequence homology to the E.coli xth (exonuclease III) gene. Using in situ hybridisation, the HAP1 gene has been localised to human chromosome 14q 11.2-12.