Preliminary investigation on the effect of dietary supplemental biotin and palm kernel oil on blood, liver and kidney lipids in chicks.

A total of 480 day-old broiler chicks were used in two trials conducted to investigate the performance and lipid contents of blood, liver and kidneys of birds when fed varying levels of palm kernel oil (0% and 2%) and biotin (40, 80, 120, 160, 200 and 240 mcg/kg feed) in a 2 x 6 factorial experimental design. The results showed that blood, liver and kidney lipid concentrations were significantly affected by dietary biotin treatments. While total lipid, free fatty acid, triglyceride and cholesterol contents were negatively correlated with dietary biotin level, phospholipid concentrations were positively correlated. Biotin-deficient chicks had significantly higher total lipid, free fatty acid, triglyceride and cholesterol but lower phospholipid contents in their blood and the two organs. Supplementation of the diet with 2% palm kernel oil significantly elevated blood phospholipid concentration, but depressed the accumulation of the other lipid fractions in both organs and the blood of birds. Blood, liver and kidney cholesterol concentrations were not affected by 2% fat supplementation. Observation on the lipid parameters coupled with the results on feed utilisation appeared to suggest that a minimum of 120 mcg of the vitamin per kilogram of diet was required by broiler chicks for optimum performance.


Sunflower meal as a major vegetable protein source in layers' ration.

An experiment was conducted with Starcross layers to evaluate the replacement value of sunflower meal (SFM) for cotton seed meal (CSM) and sesame meal (SM). The birds (aged 24 weeks) were given 4 isonitrogenous and isocaloric rations containing 0, 5, 10 and 15% dehulled sunflower meal. The substitution of SFM for CSM and SM did not generally affect the egg production, feed consumption, feed conversion nor did it have any effect on the quality of eggs as measured by Haugh Units (HU) and yolk index. Egg shell thickness, however, improved by increasing dietary levels of SFM. The feed costs per hen were generally lower for the SFM group since SFM was cheaper than CSM and SM.


[Concentrations of metabolites and hormones in the blood plasma of lactating dairy cows during treatment with recombinant bovine somatotropin (BST)].

Sixteen lactating dairy cows were submitted to six injections in a four-week interval of either 640 mg prolonged release bST or of 640 mg saline (control, n = 8). The bST-treated animals were pair fed to the control cows, which were fed according to requirements. Fourteen days after each injection, blood samples were derived and analysed for metabolites and hormones. Plasma contents averaged 0.16 mmol/l non esterified fatty acids (NEFA), 2.69 mmol/l glucose, 0.43 mmol/l beta-hydroxybutyrate, 0.024 mmol/l acetoacetate, 78.8 g/l protein, 4.6 mmol/l urea, 0.81 micrograms/l insulin, 44.4 ng/l thyroxine (T4) and 1.41 ng/l triiodothyronine (T3). The bST-application was without effect on the analysed plasma contents.


[The nitrogen metabolism in the large intestine of ruminants. 8. Metabolism of intracecally infused 15N-urea with a supplement of pectin in heifers].

Three heifers with live weights of 255, 261 and 300 kg were supplied with ileo-caecal re-entrant cannulas, jugular vena catheters and bladder catheters. The ration consisted of 4 kg maize silage and 4 kg wheat straw pellets. In a previous period 50% of the digesta flow was collected over 12 h/d on 5 consecutive days and stored in a deep-freeze. During the main period the re-entrant cannulas were disrupted and the flowing digesta was quantitatively collected. Precollected digesta and pectin were infused into the distal part of cannula hourly for about 30 hours. During the first 24 hours the digesta was also supplemented with 15N-labelled urea. The amount of pectin corresponded to about 10% of digesta dry mater. An analysis of urine, faeces, digesta and blood plasma were carried out. The application of pectin increased the 15N-incorporation in the bacterial protein of faeces from 4.7% (without supplementation in an earlier experiment) to 10.5% of the introduced 15N. The ammonia-fraction of faeces was markedly higher than the bacterial fraction. The 15N-utilization of urea by the microbes of large intestine was lower in the actual trial evident than with supplementation of starch in the anterior experiment. During the pectin administration the amount of urine increased in comparison with earlier experiments and according to the literature to about the 4.5 fold. The amount of passage of 15N at the ileum cannula (recycled 15N) was 3.8% of the 15N intake. It is the same amount as in experiments in which starch was applied.


[The measuring frequency necessary for food intake and milk yield data in feeding studies in dairy cows].

An individual feeding experiment was carried out which comprised the daily milk analysis from Monday to Friday with proportional samples from the evening and morning milkings and the daily intake of the individual feed components being ascertained. For various measuring frequencies the following calculations were made: the mean values and their standard deviations, the absolute and relative control errors and their standard deviations, the standard deviations for the measuring error and the simple linear correlations and regressions between the various measuring frequencies for the individual parameters. From the results one can conclude that, in individual feeding experiments with dairy cows, five milk analyses per week do not bring about an essential improvement of accuracy in comparison with two analyses. If the composition of the ration remains the same and the same roughage is offered ad libitum, one can project the measuring of feed intake on five days per week for that of the whole week.


Effect of feed intake and dietary vitamin A levels on sheep performance.

A study was conducted on 20 "Rahmani" male lambs of 16 kg body weight fed on normal feed level and high vitamin A level (NF-HV); normal feed level and normal vitamin A level (NF-NV); low feed level and high vitamin A level (LF-HV) or low feed level and normal vitamin A level (LF-NV) till slaughter weight of 35-40 kg. Average daily gains of lambs were 211, 148, 117 and 87 g for the NF-HV, NF-NV, LF-HV and LF-NV groups, respectively. The corresponding feed conversions were 4.3, 5.9, 5.0 and 6.4 kg DM/kg gain. Raising feed intake and vitamin A level enhanced nutrients digestibility and nitrogen balance. Rumen liquor reflected higher pH value in animals fed higher intake (NF) in the first two diets. Both feed intake and vitamin A levels enhanced volatile fatty acids and ammonia concentrations in the rumen liquor. Feed intake and vitamin A levels positively affected haemoglobin content, packed cell volume, glucose, urea, total protein, vitamin A, insulin, cholesterol, phospholipids, glutamate oxaloacetate and glutamate pyruvate transaminases and alkaline phosphatase in blood Vitamin A, insulin, cholesterol and phospholipids in blood increased gradually over the experimental period. Dressing percentages were 56.2, 49.6, 49.1 and 44.6% for the experimental groups, respectively. Liver content of glycogen and vitamin A increased with increasing feed intake and vitamin A levels. It is concluded that higher levels of vitamin A supplementation (than usually recommended) is required during fattening of lambs. Low energy and normal vitamin A levels are not recommended in lambs feeding.


[Relationship between wool yield and cyst(e)ine level in blood plasma].

Publications from Williams were the stimulation to study the relations between wool yield and cyst(e)ine concentration in blood plasma. Methodical experiments were carried out and the two methods used each provided clear results. At the same time the reversible binding of a part of the free cystine in the plasma with the plasma proteins and its effect on the analysis results might be considered. After an interpretation of the two methods their use is described taking into consideration the factors influencing the results of the analysis. It is concluded that the two methods are usable for the planned purpose.


[Cold storage of wheat. 1. Ergosterol, ochratoxin A and citrinin after inoculation with Penicillium verrucosum].

Seed wheat was inoculated without having been sterilized with an ochratoxin A and citrinin forming strain of Penicillium verrucosum and stored at moisture contents of 18, 20, 22, 24, and 26% at 10 and 4 degrees C. The production of ergosterol, a chemical indicator of fungal biomass, started within the storage time investigated (240 days). Only at 18% H2O/4 degrees C an increase of the ergosterol content was not observed. Ochratoxin A and Citrinin were not detected at 18% H2O/4 degrees C and 20% H2O/4 degrees C within 240 days (detection limit: 10 and 25 micrograms/kg, respectively). At the other combinations of moisture content and temperature the first detection of the two toxins approximately coincided with the onset of ergosterol production. With increasing moisture content and temperature the time up to the start of ergosterol production decreased, whereas the production rates of ergosterol, ochratoxin A and citrinin increased. Both toxins were produced with about the same rate during a first phase of accumulation. At 20-26% H2O there was no influence of moisture content and temperature on the relation between toxin content and the simultaneously reached ergosterol content. It is recommended that wheat highly contaminated with Penicillium verrucosum should not be stored beyond the start of ergosterol production.


[Angioproliferative processes in gangrenous limbs].

The results of the investigation of the limbs amputated because of the gangrene are presented. Soft tissues close to the gangrenous parts were examined, and in 26 of 100 samples examined angioproliferative processes with the formation of capillary foci were found. Proliferating capillaries grow into the surrounding tissues, including fibrous, muscular and adipose one. The process resembles somewhat neoplastic one: there is an infiltrative growth and mitotic activity. No similar alterations in the gangrenous tissue have been described so far.


[Is localization of B-cells in the acini a normal condition or the sign of compensatory process in insulin-dependent diabetes mellitus?].

45 pancreatic biopsies from patients with insulin-dependent diabetes mellitus (IDDM), obtained in the process of surgical treatment were studied. A different degree of the Langerhans islands and morphological changes in B-cells was noted. The presence of endocrine-like cells in the acini and among the cells of the inter-acinar duct is of great interest. The granules of these cells were insulin-positive when studied immunohistochemically this permitting one to consider them as B-cells. Further study of these cells will allow to deepen the knowledge about the mechanism of IDDM development and to improve the methods of treatment.


[Lymphangion in health and in lymphedema].

Comparative analysis of lymphangion construction in different organs and sites of human body is performed. Quantitative parameters of lymphangions (length, width, volume) are found to have age and site peculiarities. Myo-myocyte contacts of the nexus type and fascicular-reticular organizational principle of the cells underly myocyte association into functional syncytium. The valve consists of a muscleless fold and valvular bolster containing muscles. Restructuring of lymphangions in lower limb lymphedema are observed. In the course of the disease progression myocyte atrophy occurs which is followed by lymphangion wall collagenization. The alterations in lower limb lymphangions are proportional to the gravity of the disease.


[Cytogenetics of neuroblastoma and glioma].

Without having the common high specific marker, the tumours of the nervous system are, however, characterized by regular quantitative and structural changes of chromosomes. In this review molecular and genetic aspects of chromosome translocations typical for the main types of the nervous system, their role in activation of protooncogenes and their conversion to oncogenes, which is able to cause tumor transformation, are considered. Additionally, the role of chromosome translocation in the inactivation of suppressor genes is discussed.


[Modification of the cellular and vascular reactions in the inflammatory focus by luteotropic hormone].

Luteotropic hormone considerably alters the dynamics of experimental inflammation. A decrease of microvascular bed activation and the density of macrophage population are observed at early stages of the inflammation. Inhibition of fibroblast proliferative activity and acceleration of their differentiation are revealed during the period of reparation. The formation of the capsule went more intensely, but collagen synthesis was inhibited and fibrous tissue formed more slowly. The results obtained prove principal possibility of inflammation regulation by means of mediators and metabolites of the stress-limiting systems.


[Histochemical study of lesions in superficial and visceral candidiasis].

Mycotic foci were studied histochemically on various experimental models of candidiasis. NAD-H, NADP-H-diaphorase, acid phosphatase and ATPase were revealed in the fungi, the activity of these enzymes depended on the state of the fungus. Diaphorase activity in the mucous membrane epithelium falls only if it is damaged by massive invasion of pseudo-mycelium. Inhibition of the enzyme activity in the visceral foci (kidney, liver, heart) occurs only in case of pronounced destruction and is not observed at the distance from the fungi. The results do not confirm the idea of fungal secretion of mycotoxins penetrating into the surrounding tissues and damaging them.


Neutral glycolipid abnormalities in a t-complex mutant mouse embryo.

The content of neutral glycolipids was studied in normal and twl/twl mutant mouse embryos at embryonic day 11 (E-11). The twl mutation is part of the T/t complex on chromosome 17 and causes embryonic lethality from defects in the developing neural tube. Previous studies suggested that the mutation could involve a defect in ganglioside biosynthesis. Although the total neutral glycolipid content was similar in the normal and mutant whole embryos (approximately 80 nmol glucose/100 mg dry weight), marked differences were detected for the distribution of specific glycolipids. The content of lactosylceramide, globotriaosylceramide, and globotetraosylceramide was significantly higher in the mutant than in the normal embryos, whereas that of glucosylceramide was significantly reduced. The Forssman glycolipid was slightly elevated. The neutral glycolipid composition was similar in embryonic head and body regions of normal embryos, suggesting that the glycolipid abnormalities observed in the mutants are expressed in most embryonic cells and tissues. These and the previously reported ganglioside abnormalities in the twl/twl mutants could result from an inherited defect in glycolipid biosynthesis.


X-linked glucose-6-phosphate dehydrogenase (G6PD) and autosomal 6-phosphogluconate dehydrogenase (6PGD) polymorphisms in baboons.

Electrophoretic polymorphisms of glucose-6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) were examined in captive colonies of five subspecies of baboons (Papio hamadryas). Phenotype frequencies and family data verified the X-linked inheritance of the G6PD polymorphism. Insufficient family data were available to confirm autosomal inheritance of the 6PGD polymorphism, but the electrophoretic patterns of variant types (putative heterozygotes) suggested the codominant expression of alleles at an autosomal locus. Implications of the G6PD polymorphism are discussed with regard to its utility as a marker system for research on X-chromosome inactivation during baboon development and for studies of clonal cell proliferation and/or cell selection during the development of atherosclerotic lesions in the baboon model.


Antithrombin III (AT3) polymorphism in the marsupial Monodelphis domestica: identification and genetics.

Antithrombin III polymorphism was observed in the gray short-tailed opossum, Monodelphis domestica, by either one-dimensional polyacrylamide gel electrophoresis (PAGE; pH 7.9), two-dimensional PAGE (agarose, pH 5.4; 12% T, pH 7.9), or isoelectric focusing (pH 4.2-4.9) followed by immunoblotting with rabbit antiserum to human antithrombin III. Family studies demonstrated an inheritance of three codominant autosomal alleles, AT3A, AT3B, and AT3C, and a population study revealed frequencies of 0.70, 0.10, and 0.20, respectively.


Dipeptidase-C in Drosophila melanogaster: genetic, ontogenetic, and tissue-specific variation.

Dip-A, Dip-B, and Dip-C constitute structural genes for three peptidic enzymes in Drosophila melanogaster distinct from the leucine aminopeptidases. Their ontogenetic and tissue distributions of activities suggest the involvement of these enzymes in a general metabolic role, such as the regulation of amino acid and oligopeptide pools to make amino acids available for protein synthesis. Screening of chromosome substitution isogenic lines for DIP-C activity indicated that, like DIP-A and DIP-B, unlinked activity modifiers exist for Dip-C. The developmental profiles of dipeptidase activities are very similar, except in the pupal stage, during which DIP-C activity is markedly low compared to the other two enzymes. Intercorrelations of dipeptidase activities vary ontogenetically, which is consistent with the need for coordinate expression of these enzymes during certain developmental stages. Tissue-specific expression of dipeptidases in larvae and adults are also similar, although the relative levels of DIP-A activity differ from those of DIP-B and DIP-C in certain organs and body parts. Some of the differences among chromosome substitution lines for dipeptidase activities appear to be systemic, while others are developmental stage-specific and tissue-specific. Second- and third-chromosome variants for DIP-C activity differed in their tissue distribution. This is consistent with the presence of temporal and spatial variants in natural populations for other Drosophila enzymes.


Analysis of isozyme loci in the face fly, Musca autumnalis DeGeer.

Vertical polyacrylamide gel electrophoresis was used to resolve enzymes at 50 putative loci in Iowa face flies, a recently arrived, colonizing, Palearctic species. Sixty-two percent of the 50 loci were polymorphic. The mean number of alleles per polymorphic locus was 3.2 +/- 1.38; the mean among all loci was 2.38 +/- 1.62 alleles. The effective number of alleles among 50 loci was 1.4 +/- 0.62. Mean observed and expected heterozygosities for the 50 face fly loci were 0.167 +/- 0.037 and 0.186 +/- 0.031, respectively. Comparison of the electrophoretic data for Musca autumnalis and for M. domestica L. showed similar high levels of gene diversity. A survey of gene diversity at 12 loci among six geographically independent laboratory colonies demonstrated statistically significant genetic differentiation that was probably due to drift after colonization.


Molecular and biochemical elucidation of a cellular phenotype characterized by adenine analogue resistance in the presence of high levels of adenine phosphoribosyltransferase activity.

A mouse embryonal carcinoma cell line isolated for resistance to the adenine analogue 2,6-diaminopurine (DAP) was found to have near-wild-type levels of adenine phosphoribosyltransferase (APRT) activity in a cell-free assay. This DAP-resistant (DAPr) cell line, termed H29D1, also exhibited near-wild-type levels of adenine accumulation and the ability to grow in medium containing azaserine and adenine. Growth in this medium requires high levels of intracellular APRT activity. Using the polymerase chain reaction (PCR) and the dideoxy chain termination sequencing technique, an A-->G transition was discovered in exon 3 of the aprt gene in H29D1. This mutation resulted in an Arg-to-Gln change at amino acid 87 of the APRT protein that, in turn, resulted in a decreased affinity for adenine. An increased sensitivity of APRT to inhibition by AMP was observed when comparing H29D1 to P19, the parental cell line. Using a transgene containing the A-->G mutation, we demonstrated that this mutation is responsible for the biochemical and cellular phenotypes observed for the H29D1 cell line. The approach used in this study provides a definitive method for linking a mutation to a specific cellular phenotype.


Two-dimensional electrophoretic protein patterns of reciprocal hybrids of the mouse strains DBA and C57BL.

Two-dimensional electrophoretic patterns of cytoplasmic liver proteins of the mouse were investigated in reciprocal hybrids of the two inbred mouse strains DBA and C57BL in order to establish whether strain-specific protein variants reveal a mode of inheritance compatible with the concept of genomic imprinting. Variants of this type were found to account for about 11% of approximately 200 variant protein spots scrutinized. Transmission of the maternal form of a variant protein was more frequent than transmission of the paternal form. Maternal/paternal transmission was observed only for proteins showing strain variations in their amount. The results are discussed in terms of the frequency of imprinted genes.


A triangular cutting arteriotomy scissors facilitating end-to-side anastomoses.

A new microsurgical triangular cutting scissors facilitating end-to-side anastomoses has been developed. The instrument removes oval pieces of tissue from the vascular wall with a single cut. This has several advantages over conventional double cuts with straight microscissors. Preferential use of end-to-side anastomoses has been advocated by several authors, one of the main reasons for this being preservation of the distal blood flow in the artery used in the recipient bed. It also has been reported that vascular occlusions are less frequent with this type of coupling. Another important indication for end-to-side anastomoses is a significant discrepancy in vessel size. One of the more demanding tasks in making an end-to-side anastomosis is the performance of a perfect arteriotomy. With the aim of facilitating the arteriotomy in end-to-side anastomoses, a new microsurgical instrument has been developed.


Alfentanil increases intracranial pressure when intracranial compliance is low.

Intracranial pressure increases following the administration of alfentanil 1 mg are reported in five patients with suspected normal pressure hydrocephalus who were undergoing infusion of saline into a lateral ventricle to measure cerebrospinal fluid outflow resistance. The increase in intracranial pressure was accompanied by a fall in mean arterial pressure. These observations show that, when the intracranial compliance is reduced, alfentanil can cause considerable increases in intracranial pressure and decreases in cerebral perfusion pressure.


Factors associated with pediatricians' participation in Medicaid in North Carolina.

To describe the relative importance of factors influencing pediatricians' participation in Medicaid in North Carolina. Questionnaire survey. Nonacademic primary care pediatricians in direct patient care at least 50% of the time; 332 (85%) of the 389 eligible pediatricians responded. Proportion of pediatricians who restricted Medicaid patients' access to their practices. The association between restricting access and the following factors was assessed: Medicaid reimbursement, pediatricians' demographic characteristics, knowledge of the Medicaid program, attitudes toward Medicaid patients and the Medicaid program, and beliefs about whether other physicians were available to care for Medicaid patients. Twenty-nine percent of pediatricians restricted Medicaid patients' access to their practices. The proportion of pediatricians restricting access was 62% in cities, 13% in medium-sized towns, and 12% in small towns (P less than .001), but the proportion of pediatricians in cities who restricted access varied from 87% to 22%. Pediatricians who received a higher proportion of their usual fee were less likely to restrict Medicaid patients' access. The relationship between Medicaid payment and restricted access was substantially weakened after controlling for the following factors: (1) the size of the community, (2) pediatricians' attitudes toward Medicaid payment, (3) their perceptions that they were too busy to care for Medicaid patients, and (4) whether there were other resources for the care of Medicaid patients. At comparable levels of payment, rural pediatricians were about six times less likely than urban pediatricians to restrict access. Pediatricians who knew less about Medicaid reimbursement also restricted access more often. Whether or not they restricted access to new Medicaid patients, pediatricians provided acute, preventive, hospital, and emergency care to the Medicaid patients who were already in their practices. Existing resources for the care of Medicaid patients, pediatricians' economic dependence on Medicaid, and the local norms of practice may be important factors in pediatricians' decision to participate in Medicaid. Increasing reimbursement will have only modest effects on Medicaid participation. Strategies to improve participation should also address pediatricians' knowledge of the Medicaid program and enlist the support of community physicians.


Seronegative generalized myasthenia gravis: low frequency of thymic pathology.

We have reviewed thymus histology, preoperative serum acetylcholine receptor antibody status, and clinical features of all 50 patients who underwent thymectomy for generalized myasthenia gravis in the University of British Columbia-affiliated hospitals over the last 8 years. Seven patients had thymoma, 25 had hyperplasia, and 18 had a normal thymus. The seven thymoma patients all had severe limb involvement and all had circulating antibodies against the acetylcholine receptor. Patients without circulating antibodies had only minimal limb involvement and half of them had a normal thymus. Only two of the 32 (6%) patients with abnormal thymus were antibody-negative; this contrasts with six of the 18 patients (33%) with normal thymus. We conclude that seronegative patients with generalized myasthenia gravis constitute a distinct subgroup which may not be immunologically homogeneous and could be subgrouped with normal or abnormal thymic histology.


Recurrent encephalopathy and seizures in a US native with HTLV-I-associated myelopathy/tropical spastic paraparesis: a clinicopathologic study.

A patient with progressive spastic paraparesis originally ascribed to multiple sclerosis developed recurrent encephalopathy and seizures. A diagnosis of HTLV-I-associated myelopathy/tropical spastic paraparesis was established prior to death. Autopsy confirmed chronic inflammatory myelopathy and active inflammation in the white matter of the temporal lobes.


Scleral buckle with diathermy for simple retinal detachments. 100 pneumatic retinopexy eligible eyes.

One hundred consecutive pneumatic retinopexy eligible eyes underwent outpatient scleral buckle and diathermy. Follow-up averaged 31 months. Four cases required a second operation. Absence of an encircling band was the only significant risk factor (P = 0.00000028) for the three redetachments because of new or missed retinal tears. Although 18 patients had some other disease that reduced visual acuity before the retina detached, final visual acuity was 20/40 or better in 79 patients. Matched with a reported series treated by pneumatic retinopexy with cryopexy, these scleral buckle with diathermy cases exhibit significantly better final visual acuity with fewer major complications. The final acuity is also better than for two similar series, one treated with temporary balloon buckle and the second with scleral buckle and cryopexy. Use of diathermy rather than cryotherapy in the zone of retinal detachment may be the reason for the current group's better final acuity results.


Tuberculous abscesses in patients with AIDS.

Five cases of large tuberculous abscesses in patients with AIDS were observed over a 2-year period at the New York Veterans Affairs Medical Center. These cases represent 11.6% of the 43 cases of tuberculosis diagnosed in patients with AIDS during that period. The abscesses were located in the liver, abdominal wall, psoas muscle, mediastinum, and peripancreatic area. All patients presented with localized pain or swelling, and four of five patients had fever. The diagnosis was made on the basis of detection of abscesses on computed tomography (CT) and the results of culture of abscess material obtained by CT-guided aspiration. CT-guided therapeutic drainage was performed in two cases. Despite administration of therapy, two of five patients died of tuberculous infection. Formation of tuberculous abscesses appears to be a common complication of tuberculosis in patients with AIDS. This diagnosis should be considered for patients with AIDS who have fever and localized pain or swelling.


Which hormone tests for the diagnosis of polycystic ovary syndrome?

To assess the frequency of abnormal values for hormone measurements commonly used in the biochemical diagnosis of polycystic ovary syndrome (PCOS). Hormone measurements in 63 unselected women with clinical and ultrasound diagnosis of PCOS attending gynaecological and general endocrine clinics in a District General Hospital were compared with those from a group of 20 normal ovulatory controls in the early follicular phase of their cycles. Serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), LH/FSH ratio, total testosterone, derived free testosterone, sex hormone binding globulin, androstenedione, and dehydroepiandrosterone (DHEA) were measured by radioimmunoassays. LH and FSH measured by two different assays. The mean serum LH and LH/FSH ratio were significantly (P less than 0.01) higher in the women with PCOS compared with the normal group, but these two measurements were in the abnormal range for only 35% and 41-44%, respectively. Absolute gonadotrophin values were significantly different using the two assay methods, which employed the same reference preparation as standard. Mean serum total testosterone concentration was significantly higher in the PCOS group and was the most frequently (70%) abnormal biochemical marker for PCOS. Sex hormone binding globulin (SHBG) did not differ significantly between the two groups but showed a negative correlation with body mass index in women with PCOS. The combination of SHBG and testosterone to derive a free testosterone value did not further aid the biochemical diagnosis of PCOS. Androstenedione was significantly higher in the PCOS group than in the control group, with a frequency of 53%. There was no significant difference in DHEA-S between the two groups. When typical ovarian ultrasound appearances plus the clinical features of oligomenorrhea and/or hirsuitism were used to define PCOS total testosterone was the best single hormonal marker of the condition. Testosterone, androstenedione or LH, either alone or in combination, were raised in 86% of women with PCOS and these should be the definitive hormonal tests. Using LH/FSH ratio as a biochemical criterion for diagnosis of PCOS should be abandoned because of its low sensitivity. To be of value the normal range for all hormones should be precisely defined in a group of regularly ovulating women in the early follicular phase of the cycle for the assay used in each laboratory.


Hepatic arterial infusion of floxuridine in patients with liver metastases from colorectal carcinoma: long-term results of a prospective randomized trial.

A multicentric randomized study that compared patients who received intrahepatic arterial infusion (HAI) to a group of patients who did not receive HAI (control group) was performed for unresectable hepatic metastases from primary colorectal carcinoma. One hundred sixty-six patients were assigned randomly to HAI of floxuridine (5 fluoro-2'deoxyuridine [FUDR]) 0.3 mg/kg/d for 14 days every 4 weeks or to the control group; this latter group, depending on the investigator's choice, was either under observation or received systemic fluorouracil (5-FU). The same regimen of systemic 5-FU also was administered to the HAI group in the event of extrahepatic progression. No crossover from the control group to the HAI group was permitted. The mean duration of follow-up was 54 months (range, 31 to 72), and 163 patients were analyzed. A significant improvement was observed in the survival rate for the 81 patients assigned to HAI group (P less than .02) with a 1-year survival rate of 64% versus 44% in the control group (82 patients). The 2-year survival rate was 23% versus 13%. The median survival was 15 months versus 11 months for the HAI group and the control group, respectively. Survival was better for patients with a less than 30% liver involvement, and for those treated in more specialized centers. The hepatotoxic effects of HAI were observed in 47 patients (chemical hepatitis [n = 28], and biliary sclerosis [n = 19]). The 1-year rate of sclerosing cholangitis was equal to 25%. Gastrointestinal toxicity was infrequent and consisted of gastritis or diarrhea. Therapy with HAI of FUDR improves the survival of patients with liver metastases over colorectal carcinoma. However, the methods that are used to diminish the toxicity of HAI and efficient systemic chemotherapy, such as a combination of 5-FU and leucovorin, are required to prevent extrahepatic metastases.


Performance characteristics of a 511-keV collimator for imaging positron emitters with a standard gamma-camera.

Line-source experiments were conducted to assess the performance of a gamma-camera equipped with a specially designed 511-keV collimator for the planar imaging of positron emitters. The results were compared with the camera performance with routinely used collimators and radionuclides (thallium-201, technetium-99m and gallium-67). With positron emitters, scatter contributed less to the widening of the line spread function than with radionuclides emitting lower photon energies. These observations can be explained by the relative deterioration in the discrimination power of the gamma-camera to reject scattered radiation at low energies. Planar 511-keV imaging may provide relevant clinical information, as we showed by fluorodeoxyglucose studies in a patient with a myocardial infarction and in a patient with a malignant lymphoma. It is concluded that positron emitters can be effectively applied for planar imaging with the generally available gamma-cameras. This study implies that radiotracers developed for positron emission tomography may find a place in the practice of conventional nuclear medicine.


Variability in consultation rates and practitioner level of diagnostic certainty.

This study was designed to determine the variability in the consultation practices of family physicians and family nurse practitioners in an urban group practice, and to examine the relation between the referring practitioner's diagnostic certainty at the time of the consultation request, the specificity of the request to the consultant, and the frequency with which the practitioner ordered consultations in that specialty. This study examined consultations requested by six family physicians and two family nurse practitioners that were directed to nine medical and surgical specialties in connection with 35,218 family practice visits made over a 19-month period in an urban family practice. Requests for consultation were scored based on the level of certainty of the referring practitioner's diagnosis at the time of consultation and the specificity of the consultation request. There was nearly a fivefold variation in the overall specialty consultation rates among providers, with even greater variation in each individual specialty area studied. Referral rate correlated positively with certainty of diagnosis (r = .40). There was no consistent relation between request specificity and referral rate. Consultation rates vary widely, even among family physicians and nurse practitioners within the same practice. The rate at which a practitioner orders consultations in a specialty area increases in relation to the practitioner's certainty of the diagnosis at the time the consultation was requested. Diagnostic certainty is discussed as a possible indicator of a referring practitioner's knowledge in a specialty area, implying that the greater a practitioner's knowledge in a specialty area, the more he or she consults with specialists in the field. Financial incentives that attempt to decrease specialty consultation should be reexamined in light of these findings.


Mitral valve prolapse: two dimensional echocardiography reveals a high prevalence in three to twelve year old children.

The prevalence of mitral valve prolapse (MVP) appears to be age related, MVP being commoner in children as compared to adults. This suggests that asymptomatic MVP may be most frequent in children who are very young. In this study, to better define the prevalence of MVP in young children, we used two dimensional echocardiography and prospectively surveyed 213 healthy urban school children between 3 and 12 years of age. MVP was diagnosed when prolapse of mitral leaflet/s was demonstrated by both two-dimensional and M-mode echocardiography at parasternal long-axis views. Overall, MVP was found in 28 of 213 (13.1%) children. MVP was similarly prevalent in all age groups studied (3-5.9 years: 13 of 83 (13.5%); 6-8.9 years: 9 of 71 (11.2%); and 9-12 years: 6 of 31 (16.2%) children; Chi square = 0.57, p greater than 0.5). Univariate analysis showed that the prevalence of MVP was independent of sex, birth weight, resting heart rate and systolic or diastolic blood pressure. A mid systolic murmur was present in 50.6% of the children although it correlated with echocardiographic diagnosis of MVP in only 39.3%. The left ventricular size or wall thickness and mitral EF and DE slopes were similar in children either with or without MVP. Our results indicate that asymptomatic MVP is frequent in children upto 12 years of age. As a diagnostic test of MVP, presence of apical systolic murmur is considerably inferior to echocardiography. No morphological left ventricular correlates were identified in MVP.


Alternative models for specialty nursing practice.

Specialization in nursing dates from the early part of the twentieth century. Since 1925 the specialty movement has escalated, and the specialty programs have been moving to the master's degree level. There are two different models for specialization: One is the nursing model that focuses on advanced content from the behavioral sciences, education, and nursing. The specialists who use the nursing model focus on the caring role of the nurse. The specialists who follow the other model, the collaborative model, bring their nursing skills with them, and they acquire advanced knowledge of medicine and the behavioral and biological sciences. They work closely with physicians, diagnosing and treating patients. Most of the specialists within the collaborative model are covered by an additional level of state certification or licensure.


A randomized controlled trial of perioperative rifampin/trimethoprim in cerebrospinal fluid shunt surgery.

A randomized, double-blind, placebo-controlled trial of perioperative rifampin-trimethoprim was undertaken at the Hospital for Sick Children from March 1984 to October 1987, in which 243 patients undergoing 300 cerebrospinal fluid (CSF) shunting procedures were randomized into groups including treatment with rifampin/trimethoprim and placebo, and then followed for a minimum of 2 years. Patients were stratified prior to randomization into those with and those without meningo-myeloceles having first insertions of their shunts, and those having revisions. Patients could be entered into the study more than once, but always received the same treatment regimen once allocation had taken place. Among the patients receiving antibiotics there was an infection rate of 12%, versus 19% among patients receiving placebo. Among the surgical procedures, the rates were 9% and 15%, respectively. Because these rates of infection were a substantial increase over the rate of 7.5% overall for the few years prior to implementation of the study, and well over any acceptable rate of infection, the study was stopped before statistical significance was reached. However, had the study continued, and the proportions of patients becoming infected remained constant, we would have been able to achieve a statistically significant difference in rates of infection, and therefore demonstrate a benefit of rifampin/trimethoprim as prophylaxis against shunt infection. Methodological problems encountered in this and other studies of prevention of CSF shunt infection will be discussed.


Percutaneous absorption of drugs.

The skin is an evolutionary masterpiece of living tissue which is the final control unit for determining the local and systemic availability of any drug which must pass into and through it. In vivo in humans, many factors will affect the absorption of drugs. These include individual biological variation and may be influenced by race. The skin site of the body will also influence percutaneous absorption. Generally, those body parts exposed to the open environment (and to cosmetics, drugs and hazardous toxic substances) are most affected. Treating patients may involve single daily drug treatment or multiple daily administration. Finally, the body will be washed (normal daily process or when there is concern about skin decontamination) and this will influence percutaneous absorption. The vehicle of a drug will affect release of drug to skin. On skin, the interrelationships of this form of administration involve drug concentration, surface area exposed, frequency and time of exposure. These interrelationships determine percutaneous absorption. Accounting for all the drug administered is desirable in controlled studies. The bioavailability of the drug then is assessed in relationship to its efficacy and toxicity in drug development. There are methods, both quantitative and qualitative, in vitro and in vivo, for studying percutaneous absorption of drugs. Animal models are substituted for humans to determine percutaneous absorption. Each of these methods thus becomes a factor in determining percutaneous absorption because they predict absorption in humans. The relevance of these predictions to humans in vivo is of intense research interest. The most relevant determination of percutaneous absorption of a drug in humans is when the drug in its approved formulation is applied in vivo to humans in the intended clinical situation. Deviation from this scenario involves the introduction of variables which may alter percutaneous absorption.


Prevalence of Plesiomonas shigelloides among diarrhoeal patients in Bangladesh.

The incidence of Plesiomonas shigelloides among diarrhoeal patients attending the Dhaka Treatment Centre of the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B) from January through December 1987, has been reported. Using bile peptone broth (pH 8.8) as an enrichment medium and Salmonella-Shigella agar to isolate the organism, P. shigelloides was isolated from 838 (6.4%) of 13,142 patients, 523 (4.0%) of whom had no other pathogen isolated. The percentage of isolation was higher from stool (9.2%) than from rectal swab (5.9%) specimens (P < 0.005). The incidence of P. shigelloides was higher among male (64.0%) than female (36.0%) patients (P < 0.005). Isolation was highest among children less than five years. P. shigelloides was isolated most often in March (11.0%) and September (7.7%), indicating two seasonal peaks of incidence before and after the monsoons. All the strains were uniformly sensitive to chloramphenicol, furazolidon, gentamicin and trimethoprim-sulfamethoxazole. Sensitivity of the organism to ampicillin and tetracycline was 27.0% and 89.0%, respectively. This indicates that P. shigelloides may be an important agent of diarrhoea in our patient population.


Inhibition of target cell adhesion by monoclonal antibody to the 66 kD surface antigen of Entamoeba histolytica.

The adherence of Entamoeba histolytica trophozoites to target cells was studied using a monoclonal antibody to a major surface antigen of 66 kD. Preincubation of trophozoites with monoclonal antibody decreased their ability to adhere to and engulf erythrocytes and to destroy Chinese hamster ovary cells. The monoclonal antibody was specific for the 66 kD antigen, which is possibly a major participant in the adhesion that precedes phagocytosis and cytopathic effects.


An epidemiological perspective of injuries in the Northwest Territories.

This paper reviews the epidemiology of injuries in the multiethnic population of the Northwest Territories (NWT), Canada, based on mortality, hospital morbidity, and health survey data. Overall, injury mortality in the NWT has remained consistently higher than the Canadian national rate. In the NWT, an increasing trend was observed for motor vehicle accidents and homicide in both sexes, and suicide among males, while other accidents (including fires and drowning) have declined substantially among males. The age group which reported the highest suicide rate was shifted downward between the periods 1961-70 and 1981-86. Of the three major groups in the NWT, the overall injury mortality rate was comparable between Indians and Inuit, and their rates were twice as high as that of non-Natives. The age-standardized hospitalization rate for all injuries for the Keewatin Region was similar to that of Canada as a whole whereas the rate for the entire NWT was about twice as high. In the NWT, age-specific hospitalization rate was highest in both sexes among the elderly (age 65+), followed by those in age group 15-24. A survey of a random sample of Keewatin Region residents indicated a low usage rate for seatbelts and helmets while a significant minority of drivers (18%) admitted to drinking and driving sometimes. Such data contribute to a more comprehensive assessment of the burden of injury mortality and morbidity, an important prerequisite for the investigation of risk factors and design of intervention programs.


Septal neurons related with hippocampus increase their firing rate after long-term clomipramine.

In the rat, long-term clomipramine increases the firing rate in lateral septal neurons. Although the hippocampus is the main afference for septal nuclei, it is unknown whether clomipramine increases the firing rate in most of hippocampal-septal neurons. Therefore, the present study explored the actions of long-term clomipramine in lateral septal neurons identified by their relation to the hippocampus. In most recordings, hippocampal stimulation produced a brief excitatory short-latency response, followed by a period of inhibition of firing. These neurons increased their firing rate after clomipramine treatment. Other septal neurons not respondent to hippocampal stimulation did not respond to clomipramine treatment, either. We concluded that only hippocampal-septal neurons are clomipramine responders too, and the drug-induced enhancement of firing rate is likely to be mediated by an interneuron-mediated disinhibition process.


Effects of nimodipine on multiunit activity of several brain structures following acute global cerebral ischemia-anoxia in cats.

The effects of nimodipine, a 1,4-dihydropyridine calcium channel blocker, on multiunit activity (MUA) of several brain structures were investigated in cats during 6 h immediately following acute global cerebral ischemia-anoxia induced by a 10 min cardiorespiratory arrest (CRA), as well as in cats exposed to sham procedures corresponding to CRA. Four groups of cats were studied: 1) CRA and continuous administration of nimodipine, 1 microgram/kg/min iv during 6 h; 2) CRA and continuous administration of vehicle; 3) sham and continuous administration of nimodipine as in group 1; 4) sham and vehicle as in group 2. MUA and electroencephalogram disappeared during ischemia-anoxia; their progressive recovery occurred throughout the hours following CRA, although 6 h after CRA MUA was still lower than its control prearrest values in all the recorded subcortical structures. Delta-like waves, isolated spikes, and bursts of fast EEG waves occurred during the recovery of EEG activity. Nimodipine inhibited the otherwise increasing MUA in mesencephalic reticular formation, hippocampus and putamen, but not in ventromedial hypothalamus, during the hours following acute global cerebral ischemia-anoxia. Absence of isolated spikes and bursts of fast EEG activity was noted in the EEG of CRA-, nimodipine-treated cats. Nimodipine significantly reduced MUA in hippocampus but not in other cerebral structures in cats of the sham treated group. The results suggest the involvement of 1,4 dihydropyridine sensitive calcium channels in the cellular mechanisms related to neuronal activity after cerebral ischemia-anoxia, and the possible relationship between the effects of nimodipine on MUA and better functional conditions of the central nervous system after acute global cerebral ischemia-anoxia.


The importance of employing homologous serum free of thyronines in radioimmunoassays to assess circulating thyroid hormones in rainbow trout.

Although abundant, available information regarding the values of circulating thyronines in the rainbow trout is misleading and confusing. The present study demonstrates the importance of maintaining species serum-homology when measuring thyronines with RIA to avoid spurious results. The utilization of homologous (trout) serum free of thyronines in the standard curve allowed the standardization of accurate and specific radioimmunoassays to quantitate circulating T3 and T4 levels in 10 microliters of rainbow trout serum. Detection limit was 50 pg for both thyronines. Inter- and intra-assay coefficients of variation were minor to 10%. Mean values for T3 and T4 +/- SEM in the trout population studied (n = 34) were 23.6 +/- 1.2 and 12.0 +/- 2.3 ng/ml respectively. A positive and significant correlation (p < 0.005) between body weight and T3 was observed, confirming the active participation of this thyronine in growth processes.


Cortical and septal responses to dorsal raphe nucleus stimulation in the rat: long-term clomipramine actions.

In cerebral cortex and lateral septal nuclei different serotonergic receptor subtypes coexist, thus a different action on neuronal firing may be expected depending on the receptor activated. Dorsal raphe nucleus stimulation produced an increased rate of firing in cortical layer V, and in lateral septal nuclei. However, firing rate in cortical layer VI remained unchanged after stimulating the dorsal raphe nucleus. Clomipramine is a tricyclic which exerts its main actions on serotonergic receptors, and long-term treatment with this antidepressant produced a selective increased firing rate in lateral septal neurons, but not in cortical neurons. From an electrophysiological point of view, it is concluded that the excitatory actions on firing rate elicited by dorsal raphe nucleus stimulation or clomipramine treatment are mediated by 5-HT2 receptor subtype activation which is likely to be acting as a 5-HT1A modulator in such places where both receptor subtypes coexist.


Rapidly recurring cortical seizures induce changes in neuronal responsiveness to acetylcholine.

The majority of mechanisms proposed to explain epileptic discharge suggest an excessive synaptic input into the cell or possible changes in cellular excitability which result in a decreased firing threshold and in the presence of self-sustained activity. It is likely that these changes are caused by modifications in the membrane receptor sensitivity to a specific neurotransmitter. In view of the above, the purpose of the present study has been to evaluate the sensitivity of the postsynaptic receptor by means of the microiontophoretic applications of substances whose pharmacological effect is known, thus determining its possible involvement in the epileptic process. Changes in cortical excitability were induced by electric stimuli in the sensorimotor cortex of rats anesthetized with urethane (1 g/kg intraperitoneally), immobilized with pancuronium bromide and kept alive with mechanical respiration. The electric stimuli consisted of trains of biphasic pulses, each lasting one millisecond, with a frequency of 100 pps and with a train duration of 1 second. The response of the neuron to acetylcholine was evaluated before and after the kindling had been established. The dosage was measured in nanoamperes of microiontophoretic ejecting current. Extracellular field potentials were recorded with the central barrel of 4-barrel micropipettes. Peripheral barrels were used for iontophoretic applications of Acetylcholine (Ach .1, 1M), Atropine (25mM). One of these barrels containing NaCl (2M) was used for the automatic passage of balancing current.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of continuous, long-term administrations of acetylsalicylic acid on hematological and hemocoagulation changes in the rat.

Acetylsalicylic acid is currently being widely used either itself or in combination with the other substances for the prevention of thromboembolic events. Its effect is particularly given by the inhibition of the cyclooxygenase enzyme participating in the metabolism of eicosanoids. The recommended doses of acetylsalicylic acid as well as frequency of administrations are very different. The present work is aimed at estimating the effect of the continuous administration of acetylsalicylic acid for 28 days on hematological and hemocoagulation changes in the rat. The experimental results suggest that long-term administrations of acetylsalicylic acid should be used cautiously in human medicine. They signalize the need of long-term, aimed clinical studies of hematological parameters of patients preventively treated with acetylsalicylic acid, also with respect to the producer of this substance. It is also impossible to neglect regular controls of necessary parameters for the estimation of the efficiency of the given therapy in each patient with respect to individual reactions. The dosage should also be adjusted with respect to the age, sex and individual sensitivity of the organism.


[The bioavailability of nicorandil after oral administration of a single dose of 10 or 20 mg preparations].

To evaluate the bioavailability of 4 different formulations of nicorandil, 10 mg and 20 mg (Bracco) versus 10 mg ang 20 mg (Merck), we have carried out a study involving 24 young healthy volunteers. The drug was administered in single oral dose and the plasma concentration levels were evaluated by HPLC method. From the results obtained, we have seen that both the 2 products of 10 mg (A and B) and the 2 products of 20 mg (C and D) are equivalent and have high relative bioavailability: it reaches the values of 96.5% (A versus B) and 120% (C versus D). As regards the 2 formulation of 20 mg, the results of bioavailability are confirmed by data obtained from dissolution study in vitro, where the dissolution is almost complete in 30 minutes.


[A new cold sterilization chemical. I. Chemical characterization and mechanism of action].

The equilibria relative to a cold sterilizer and their interactions were studied to verify how many active compounds were present in solution. Microbiological activity derived from chlorine, hydrogen peroxide and peroxymonosulfate which could react also with chloride ions present in biological fluids, developing active chlorine.


Induction of endoreduplication in double minutes of the human neuroblastoma cells and the replication pattern.

Induction of endoreduplication (ERD) using Hoechst 33258 as well as colcemid was carried out in cultured neuroblastoma (NB) line cells. In these endoreduplicated cells, the majority of double minutes (DMs) appeared to take a diplochromosome like configuration to form a cluster consisting of four minute elements, assuming a complex DM. Sister chromatid differential staining (SCD) using 5-bromo-2'-deoxyuridine (BrdUrd) revealed the non-random distribution of the stained chromatids among four chromatids composing each diplochromosome, suggesting the occurrence of so-called "outside replication" of DNA strands during the process of ERD. The same pattern of differential staining was also found in the quadruple minutes of each endoreduplicated DM. Since DMs are acentric, the present results suggest that centromeres do not play any essential role in the formation of diplochromosomes observed in the conventional cytologic preparations and that centromeres are probably not responsible for the phenomenon of the "outside replication" of DNA strands.


Analysis of protocol variations on DNA yield.

A modified alkaline lysis method for preparing plasmid DNA from bacterial cells has been developed for automation implementation. The objective of this study was to develop a simplified centrifuge-based protocol that can process samples in a microwell plate. These manual experiments and parametric studies show that the alkaline lysis method can be modified significantly to enable DNA preparations to be done rapidly and reliably by an automated system. The conclusions of this study were (a) centrifugation at < 1500 g is sufficient, (b) centrifugation times need not be extended to compensate for the reduced force, (c) reactions can be done at room temperature, (d) reagent volumes can be reduced over those typically used, and (e) certain reagents can be combined to simplify the handling of fluids.


High-throughput DNA preparation system.

A system demonstrating the feasibility of high-throughput, centrifugation-based DNA separations and purifications has been constructed and tested. Samples are currently processed at a rate of 96 in approximately 2-3 h. The device implements an automation-optimized alkaline lysis protocol for the rapid extraction of plasmid or cosmid DNA from 1-ml bacteria cultures. The conditions for optimal culturing in deep-well (96 x 1 ml) microwell plates have been developed, and all sample manipulations are done within these plates. The use of microwell plates was essential to obtain high throughput and make manipulations following the DNA preparation (prep) easier because they can then be manipulated using a variety of commercially available robots. The entire prep system is constructed above a Beckman GPR centrifuge and operated under Macintosh IIcx control. This device has systems for fluid handling, microwell-plate manipulations, and centrifuge rotor alignment.


[Preliminary clinical trial of intrathecal rt-PA (TD-2061) for the prevention of cerebral vasospasm in patients with aneurysmal subarachnoid hemorrhage].

The results of preliminary clinical trial (a multicenter, open-label, dose escalation study) of intrathecal recombinant tissue plasminogen activator (rt-PA) for the prevention of cerebral vasospasm were reported. Seventeen patients admitted within 48 hours of subarachnoid hemorrhage (SAH) were enrolled in this study. Patients ranged from 42 to 69 years of age. All cases enrolled were classified in clinical grade II, III or IV according to the classification of Hunt and Kosnik and in group 3 or 4 according to Fisher's CT grading scale. Surgery for clipping the aneurysms were performed and a small silicone catheter was left in the subarachnoid space. Twenty four hours after the surgery intrathecal bolus infusion of rt-PA was started through the silicone catheter at 6-hour intervals for 3 days. Patients were divided into 4 groups based on the dosage of rt-PA for each infusion. The dosage of rt-PA for each infusion and a number of cases in each group was as follows; 25 KIU in 4 cases, 75 KIU in 6 cases, 200 KIU in 4 cases and 600 KIU in 3 cases. There was no significant difference in the clearance of subarachnoid clots between four groups. However, the occurrence of both symptomatic and angiographic vasospasm was less in the 75 KIU group than in other three groups. Intracranial bleeding complications were noted in 4 patients (1/6 in the 75 KIU group, 2/4 in the 200 KIU group and 1/3 in the 600 KIU group). Serial coagulation studies demonstrated no evidence of systemic fibrinolysis. Disorientation was noted in 2 out of 3 patients of the 600 KIU group.(ABSTRACT TRUNCATED AT 250 WORDS)


[Correlation between clinical symptoms and draining pathways in dural arteriovenous malformation of the cavernous region].

The clinical picture of dural arteriovenous malformations cannot be explained on the basis of degree of arteriovenous shunting in many cases. In this study, therefore we focused on the relation between clinical symptoms and draining pathways. Eighteen patients with dural arteriovenous malformation in cavernous region were studied before treatment. All patients underwent examination for their clinical symptoms, opthalmological examination and angiography on admission. Angiograms were used to determine the degree of arteriovenous shunting and the direction of draining pathway. The relationship between severity and shunt volume was inexplicable in many cases. However, close correlations appeared to exist between ocular hypertension and drainage pathways toward the orbit, between cranial nerve signs and drainage pathways toward the posterior fossa, and between headache and drainage pathways toward the cortical veins.


Detection of single-base mutations in a mixed population of cells: a comparison of SSCP and direct sequencing.

Single-base-pair changes in several genes have been shown to be important in carcinogenesis. We have compared the sensitivity of two commonly used techniques for detection of single-base-pair changes. Defined populations of cells were prepared by mixing wild-type Chinese hamster ovary cells with cells carrying a known mutation in the CTP synthetase gene. RNA was extracted and analyzed for the mutation by single-strand conformational polymorphism and direct sequence analysis of polymerase chain reaction products. We found that both techniques were able to detect the mutation when it was present in 25% of the cells, but that direct sequencing was slightly more sensitive and able to detect the mutation when it was present in only 20% of the population.


Histochemical techniques for locating Escherichia coli beta-galactosidase activity in transgenic organisms.

Escherichia coli beta-galactosidase is a commonly used reporter molecule for analyzing gene expression. Recently, beta-galactosidase fusions have been applied to a variety of eukaryotic systems. The techniques for constructing and introducing beta-galactosidase fusion constructs as well as soluble assays for total enzyme function have been described in detail elsewhere. This article describes histochemical techniques for analyzing organisms that contain a functional beta-galactosidase fusion construct. The object is to determine semiquantitatively which cells are expressing the beta-galactosidase fusion protein, as well as the subcellular localization of the protein. Due to its prevalence in the author's laboratory, Caenorhabditis elegans is used as a canonical organism for the detailed methods described.


[Effects of isosorbide dinitrate on regional cerebral blood flow and intracranial pressure in cats].

The effects of isosorbide dinitrate (ISDN) on regional cerebral blood flow (rCBF) and intracranial pressure (ICP) were examined in cats. A low dose of ISDN (2.5 micrograms/kg/min) infusion did not show any changes in cerebral hemodynamics. During high dose of ISDN (5.0 micrograms/kg/min) or NTP (5.0 micrograms/kg/min) infusion, mean blood pressure (mBP) decreased by 10 to 20% accompanied by decreased cerebral perfusion pressure (CPP: mBP-ICP), however, rCBF or ICP did not change. It is concluded that intravenous administrations of ISDN in a dose of 2.5-5.0 micrograms/kg/min that produce slight decrease in blood pressure did not influence on cerebral hemodynamics.


[A case of Meige's syndrome associated with post head trauma].

The pathogenesis of Meige's syndrome (MS) is controversial and has yet to be determined up to today. We studied a case of MS associated with post head trauma. The patient was a 52-year-old female. At the age of 46, she began to suffer from oro-lingual dystonia after head trauma induced by a traffic accident and the brief administration of neuroleptics to the delusion deteriorated the dystonia. She showed a wry appearance after 1 year and 6 months of the trauma and began to exhibit blepharospasms, oro-mandibular dystonia and cervical dystonia after 2 years and 3 months. For these symptoms her daily life became difficult. These symptoms were resistant to various drug therapies, although trihexyphenidyl relieved the symptoms transiently. Laboratory examinations and cranial MRI findings were normal. By surface electromyogram of ocular orbicular muscles, bilateral continuous discharge was observed. This patient was diagnosed as MS by clinical symptoms and surface electromyogram findings. It was inferred that the head trauma was associated with the development of MS. We discussed the pathogenesis of MS in the present case and it was speculated that MS was presented by a minute lesion of the brain stem which was produced at the time of the head trauma.


[Aspergillosis involving the thoracic spinal cord--an autopsy case].

A 63 year old male complained of persistent backache and productive cough. The chest X-ray revealed the fungus ball at the left apical-posterior segment and Aspergillus fumigatus was cultured from the sputum. He was treated on fulconazole and miconazole. Six months later, motor and sensory paralysis below the mamillary level and urinary and stool incontinence developed. A magnetic resonance image disclosed the destruction of the second thoracic spinal vertebra involved by the cavitated fungus ball of the left lung. Continuous peroral administration of antifungal drugs was not successful, and he expired with severe dyspnea. The autopsy revealed an extensive granulomatous and purulent change of the epidural and subdural spaces of the second to fifth thoracic spinal cord. Subdural inflammation extended to the lower thoracic and lower cervical level. Thoracic spinal cord revealed an extensive myelomalacia predominantly involving the left lateral white column, and also anterior and posterior columns. Small areas of the white matter were cystic. The left anterior horn cells revealed severe central chromatolysis. Moderate lymphocytic and plasma cell infiltration was found around the vessels within the cord. A few thrombi were found in the vein near the anterior nerve root. Central nervous system involvement of pulmonary aspergillosis is quite uncommon. However, there are a few reports of patients with paraplegia secondary to the spinal extension by aspergillus infection. Sheth et al. described that epidural and subdural granulomatous change with aspergillus abscesses and spinal cord myelomalacia is comparable to metastatic carcinoma. However, the aspergillus infection in the spinal cord is more extensive and destructive.(ABSTRACT TRUNCATED AT 250 WORDS)


[Intracranial primary malignant lymphoma following Behçet's disease--case report].

We reported a case of intracranial primary malignant lymphoma following Behçet disease treated with colchicine. A 43-year-old female with a past history of oral ulcer and folliculitis visited the Department of Ophthalmology on December, 1990, because of her impaired visual acuity. A diagnosis of uveitis due to incomplete type Behçet disease was made from funduscopic examination and her past history. She had been received 1.0mg/day colchicine for six months in the outpatient clinic. She was referred to our department on August 19, 1991, with nausea and headache. On admission, neurological examination revealed slight right cerebellar ataxia showing dysmetria and dysdiadochokinesis. Computed tomography scan and magnetic resonance images demonstrated a malignant lymphoma in the left cerebellar hemisphere. Stereotaxic biopsy resulted in the B cell type malignant lymphoma (Diffuse large cell type). Galliumscintigraphy, physical examination and peripheral blood examination disclosed no systemic abnormalities. There was a reduction of lymphocyte's response to PHA (phytohemagglutinin) stimulation (1.24). The tuberculin reaction was negative. Colchicine was discontinued and the therapy with radiation and prednisolone (20 mg/day) was started under the diagnosis of intracranial primary malignant lymphoma. She was discharged without any neurological deficits on November 25, 1991. She still suffers from impaired visual acuity, but has no problems in daily life. To our knowledge no intracranial primary malignant lymphoma following Behçet disease has been reported previously. The relationship between Behçet disease treated with colchicine and the occurrence of malignant lymphoma is discussed. In our case we speculated that malignant lymphoma might have occurred in an immunosuppressive state due to administration colchicine.


[Epithelial cyst occurring on the anterior surface of the brain stem--case report and review of the literatures].

Epithelial cysts in the posterior fossa are very rare, and only 22 cases have been reported in literatures. We report a case of epithelial cyst occurring on the anterior surface of the brain stem. A 31-year-old man was admitted to our hospital complaining of right hemiparesis and dysarthria. Plain CT showed a heterogeneous (isodense-hyperdense) mass in front of the pons, and was not enhanced after injection of contrast medium. As for findings of MRI, the tumor revealed as heterogeneous (isointense-hyperintense) mass on T1-weighted image and also heterogeneous (hypointense-hyperintense) mass on T2-weighted image. We removed partially the tumor which content was like yogurt. The histological findings of the cyst wall revealed a pseudostratified columnar ciliated epithelium.


[A case of frontal lobe epilepsy presenting with recurrent speech arrest].

We report a 55-year-old right-handed man with frontal lobe epilepsy manifesting recurrent speech arrest. He was known to have hypertension, hypertriglyceridemia, and gout. In the three days prior to admission, he had episodes of sudden inability to talk. These episodes lasted 10 to 30 seconds and recurred ten to twenty times a day. On admission, speech comprehension and other mental functions were normal, as were findings on neurologic examination. During the period of speech arrest, he understood spoken commands, and there was no abnormal motor activity or paresis. The episodes of speech arrest were thought to be short aphasic periods due to transient ischemic attacks in the left carotid territory. Computed tomography and magnetic resonance imaging demonstrated a small calcified lesion in the upper medial portion of the left frontal lobe. Left internal carotid angiography demonstrated no abnormal findings. After neuroradiological examination finished, he suddenly raised his right hand and followed it with his gaze and a right head turn. The EEG seizure pattern in which 20-25 Hz activity began in the left fronto-central region and spread rapidly to the right fronto-central region, which after about 8 seconds was replaced by 12-14 Hz flattening rhythmic polyspikes was detected 9 times within 60 minutes. It is most unusual for supplementary motor area seizure to present pure paroxysmal speech arrest without accompanying paroxysmal motor activity. As in our case, epileptic arrest of speech may be confused with a transient ischemic attack of the dominant hemisphere.


[High-dose intravenous immunoglobulin treatment in chronic inflammatory demyelinating polyneuropathy].

We treated two patients with chronic inflammatory demyelinating polyneuropathy (CIDP) with high-dose intravenous immunoglobulin (HIG). The patients received 400 mg/kg of immunoglobulin a day for five days. One patient, who had failed to respond to prednisolone before, was treated with HIG, 18 months after the onset. His motor symptoms resolved immediately after the commencement of HIG. Electrophysiologically, the compound muscle action potentials increased in amplitude in all nerves examined and F wave reappeared in the left median nerve. The electrophysiological changes were compatible with improvement of conduction blocks. This patient had headache and exanthema during the HIG therapy, but they settled after cessation of the infusion. The other patient was administered HIG as an initial treatment, four months after the onset. HIG was of no effect in this case, but he showed remarkable recovery during the following prednisolone therapy. Although corticosteroid therapy is the first choice for CIDP, there are CIDP patients who do not respond to steroid or can not complete the steroid therapy because of adverse effects. HIG is an expectative and recommendable treatment for the steroid resistant CIDP patients.


[Surgical treatment of brain stem lesions with reference to cavernous angioma].

The surgical indications for localized brain stem lesions were evaluated retrospectively through the clinical results of 14 patients: 5 cavernous angiomas and 9 gliomas. Cavernous angiomas were located in fourth ventricle floor (2 cases), in dorsal midbrain (1 case), in right cerebellar peduncle (1 case), and in medulla oblongata (1 case). Those cases had direct surgery because of relapse of clinical symptoms and enlargement of the lesions on follow-up MR imagings. Each lesion was extirpated totally. Consequently, the majority of neurological deficits before operation improved. Therefore, radical extirpation in brain stem cavernous angioma was strongly recommended. Also, total, subtotal resection was performed for gliomas localized in brain stem: 2 low grade astrocytomas, 3 malignant astrocytomas, 3 plexus papillomas, and 1 ependymoma. Most of cases improved without new neurological deficits after surgery. In addition, MR imaging was considered to be essential to accurate diagnosis and surgical strategies for brain stem lesions.


[Serotonergic mechanisms in hippocampal kindled seizures--inhibitory effects of L-5-hydroxytryptophan and 8-hydroxy-2-(di-n-propylamino)tetralin].

Serotonin (5-HT) has been considered to possess an inhibitory action against the kindling development, but the role of 5-HT in kindled seizures is unclear. Furthermore, most previous studies have dealt with amygdaloid kindling. To clarify the role of the 5-HT system in epilepsy, we examined the effects of 5-hydroxytryptophan (5-HTP), a precursor which elevates brain 5-HT, and 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), a 5-HT 1 a receptor agonist, on seizures kindled from the feline hippocampus (HIP). Following the completion of HIP kindling, five cats received 0.9% saline, 5-HTP (20 or 40 mg/kg, i.p.) or 8-OH-DPAT (0.1 or 1.0 mg/kg, i.v.). Drugs were administered 15 min (8-OH-DPAT) or 1 hr (5-HTP) prior to electrical stimulation at the generalized seizure triggering threshold, and the anticonvulsant effects were assessed by the behavioral seizure stage and afterdischarge (AD) duration. Both 5-HTP and 8-OH-DPAT suppressed dose-dependently HIP-kindled seizures. The administration of 5-HTP at 40 mg/kg and of 8-OH-DPAT at 1.0 mg/kg produced a marked or complete suppression of HIP-kindled seizures in most of the cats tested, and was found to significantly reduce the seizure stage when compared with the saline control. Both drugs tended to shorten the AD duration, but this effect did not reach statistically significant levels. The present data suggest that the 5-HT system plays an important role in HIP-kindled seizures, and that the 5-HT 1 a receptors have an inhibitory effect on the kindled focal epileptic activity of the HIP.


[Local cerebral blood flow and its CO2 reactivity in the ischemic cerebrovascular diseases before and after bypass].

We evaluated the effects of the STA-MCA anastomosis on local cerebral blood flow (LCBF) and CO2 cerebrovascular reactivity (CVR) in ischemic cerebrovascular diseases (CVD). [Patients and Methods] We examined the LCBF and CVR of 21 patients with ischemic CVD subjected standard STA-MCA anastomosis. During surgery, LCBF was continuously measured using the thermal diffusion method sometimes in a state of decreased PaCO2 (median: 7.2mmHg) before and after bypass. CVR was represented as % delta LCBF/delta CO2. [Results] 1) In the control group (with non-ruptured aneurysm, n = 7), LCBF was 58.6 +/- 14.1 ml/100g/min and % delta LCBF/delta CO2 was 4.68 +/- 1.68%/mmHg. 2) Before bypass, LCBF was 46.7 +/- 10.3ml/100g/min. It was significantly (p < 0.05) lower than control level. After bypass, LCBF increased significantly (p < 0.05) to 62.0 +/- 14.3 ml/100g/min. 3) Before bypass, % delta LCBF/delta CO2 was -1.36 +/- 4.34%/mmHg. In the 14 cases (66.7%), % delta LCBF/delta CO2 was below the 0%/mmHg (inverse steal effect). 4) After bypass, it increased to 0.08 +/- 2.73%/mmHg. 11 cases (52.4%) showed % delta LCBF/delta CO2 was above the 0%/mmHg. 5) In the group with a preoperative % delta LCBF/delta CO2 of less than 0%/mmHg, preoperative LCBF was 43.6 +/- 9.3 ml/100g/min, which was significantly low (p < 0.01) compared to that in the group with higher % delta LCBF/delta CO2. After bypass, LCBF and % delta LCBF/delta CO2 remarkably increased in the group with % delta LCBF/delta CO2 below 0%/mmHg. [Conclusion] Before bypass, LCBF was low and CVR was severely disturbed. In the 14 cases % delta LCBF/delta CO2 was below 0%/mmHg.(ABSTRACT TRUNCATED AT 250 WORDS)


[The analysis of nuclear organizer regions of astrocytomas with various histologic malignancies].

Nucleolar organizer regions (NORs) correspond to the loops of DNA which encode the ribosomal RNA. Acid proteins related to NORs can be stained by the silver colloidal technique (AgNORs). Since the configurations of AgNORs may be related to the protein metabolism or the proliferative activity of the cell, we tried to evaluate the corelationship between the morphology of AgNOR and the histologic malignancy in astrocytic tumors. For the quantitative evaluation the histographic pattern of AgNORs was analysed. Twenty-seven surgical specimens of astrocytomas (astrocytoma; 7 Cases, anaplastic astrocytoma; 9 cases, glioblastoma; 11 cases) were examined. The average of the means of AgNOR count in astrocytoma, anaplastic astrocytoma, glioblastoma were 1.68, 1.85 and 2.76 respectively. The averages of standard deviations (S. D.) of AgNOR count were 0.87, 1.03 and 1.26, respectively. In those tumors, the AgNOR histograms were flattered and the means and S. D. increased significantly as the malignancy increased. We speculate that the increased number and variations of AgNOR count could be a reflection of phenotypic alterations of astrocytoma cells such as cellular anaplasia and pleomorphism.


[Electrophysiological evaluation of human consciousness level by "Automated Fluctuation Analysis" of human high frequency EEG].

This study is done to clarify the clinical meaning of "Automated Fluctuation Analysis" of high frequency EEG in man especially focused on the fine alteration of consciousness level of the subjects. Twenty normal volunteers were utilized for this study. They were divided into two groups, the subjects who felt sleepiness (Group S; N = 8) and the subjects who denied it (Group W; N = 12), during the EEG recording. "Automated Fluctuation Analysis" of high frequency EEG had been presented in our previous paper. In short, it is made of three steps, amplification of EEG signal, A/D conversion and Fast Fourier Transform by signal processor and extraction of Lorentzian parameters. Power spectral density (PSD) was displayed on log-log graph. Then the third step is performed by the best curve fitting program to the following equation, S (f) = S1/[1 + (f/fc1)2] + S2/ [1 + (f/fc2)2], where S(f) is power spectral density at any frequency f, S1 and S2 are plateau level of initial and second Lorentz, respectively and fc1 and fc2 are the corner or half power frequency of initial and second Lorentz, respectively. The algorithm of this program to extract these parameters were mathematically based on Brown & Dennis. As results, 1. PSD of human high frequency EEG was composed of double Lorentzians and vanished into white level within 1kHz. 2. A topographical display of S1 value revealed hyperfrontal in group W, which is in accordance with the cerebral blood flow study by Ingvar.(ABSTRACT TRUNCATED AT 250 WORDS)


[Nicergoline, an ergot alkaloid, improves ischemic brain damage by ameliorating the decreased cerebral blood flow and metabolism in spontaneously hypertensive rats].

Effects of ergot alkaloids, nicergoline (NIC), on survival rate, brain water content, local cerebral blood flow (LCBF: 14C-iodoantipyrine) and glucose utilization (LCGU: 14C-2-deoxyglucose) were examined after bilateral carotid artery occlusion (BCAO) in spontaneously hypertensive rats. Two series of study were performed; the permanent BCAO and 3-hr-BCAO study. After permanent BCAO, the survival rate at 24 hrs of 32% (8 mg/kg, i.p.) or 38% (16mg/kg) in NIC group was higher than that in non-treated group (12%). At the end of 3-hr-BCAO, the increase in water content (dry-wet) in di-mesencephalon was less in NIC (100 micrograms/kg/min, i.v.) group than that in non-treated group. The decrease in LCBF in caudate-putamen (CP), parietal cortex (PC), thalamus (TH), hypothalamus (HT), and substantia nigra (SN) were less in NIC group than those in non-treated group. At the 2-hr-reperfusion after 3-hr-BCAO, the decrease in LCBF in TH and HT were less in NIC group than those in non-treated group. The LCGU in sensory motor cortex, CP, PC, HT, inferior colliculus and pons-reticular were higher in NIC group than those in non-treated group. From these results, it is concluded that nicergoline may have ameliorative effects on survival rate related to the prevention of decreased cerebral blood flow and metabolism following brain ischemia.


[Changes in seizure-triggering threshold and after discharge duration in seizure susceptibility].

Epilepsy is analysed on terms of two different conditions, seizure susceptibility and seizure induction. Reports on detailed changes of seizure-triggering threshold and after discharge duration in seizure susceptibility are few. We examined staged changes of seizure-triggering threshold and after discharge duration in seizure susceptibility in a cat amygdaloid kindling model. Ten crossed adult cats were used. We investigated after discharge threshold (ADT), partial seizure threshold (PST) in stage 4 group (P) and generalized seizure threshold (GST) in stage 6 group (C). At the same time, we examined after discharge duration on electroencephalograph (EEG) in each stage. Seizure-triggering threshold decreases significantly step by step. After discharge duration increases significantly step by step. It is concluded that staged decrease of seizure-triggering threshold and increase of seizure's grade and duration was seen in seizure susceptibility.


[Application of immunohistochemistry to the dead brain].

To evaluate the usefulness of immunohistochemistry for studies of brain tissues taken from patients with a clinically established diagnosis of brain death, we examined 8 brains that had been removed and fixed in formalin between 13 h and 5 days after diagnosis of brain death (Table). As a control we also examined the brain of a patient who had suffered extensive infarction with a 28-day clinical course and died of pneumonia. Routine examination of each of the brains revealed old to recent tissue necrosis associated with macrophages, capillary proliferation and/or astrocytic gliosis. The streptavidin-biotin immunoperoxidase method was applied to the materials using four monoclonal antibodies: antiglial fibrillary acidic protein (GFAP), anti-vimentin (vimentin), LN-1 and LN-3. GFAP was stained in reactive astrocytes even 5 days after diagnosis of brain death (Fig. 3, 7). Vimentin (Fig. 2, 4) was stained in reactive astrocytes until 3 days after diagnosis of brain death. LN-1 (Fig. 5) produced positive staining in resting microglias until 3 days after diagnosis of brain death. LN-3 (Fig. 1, 6, 8), which detects human leukocyte antigen-DR, stained reactive microglias and macrophages even 5 days after diagnosis of brain death. From our findings, it is considered that immunohistochemical examination is useful even in brains from patients diagnosed as brain-dead.


[Experimental syringomyelia in rabbits and rats after localized spinal arachnoiditis].

In order to produce syringomyelia, localized arachnoiditis was created in adult New Zealand albino rabbits and Wistar rats by the injection of kaolin into the thoracic spinal subarachnoid space and incision of the dura mater of the thoracic spinal cord. The rabbits and rats were divided into 3 groups; the control group, dural incision group (DG) and kaolin injection group (KG). Each rabbit was sacrificed at 4, 8, 12 and 16 weeks after the operation. Each rat was sacrificed at 8 and 16 weeks after the operation. Cavity formation in the cord of all rabbits was examined by ultrasound. All animals were perfused with 10% neutral beffered formalin at 150 cm H2O pressure, and histological examination was performed with Luxol fast blue (LFB) and hematoxylin and eosin (H&E) stains. Results obtained: (1) Cavity formation was noted in 6 of 16 DG of rabbit (37.5%), 5 of 16 KG of rabbit (31.2%) and 2 of 9 KG of rat (22.2%) with histological verification. With use of ultrasound, cavity was noted in 3 of 16 DG rabbits (12.5%) and 2 of 16 KG rabbits (18.8%). (2) Cavity formation was present in the cord adjacent to the marked adhesive arachnoiditis both in rabbits and in rats. (3) Cavity was noted in the ischemic area. (4) In 2 rabbits in which kaolin encircled whole surface of the spinal cord, hydromyelia was formed communicating with enlarged central canal caudad from the kaolin subarachnoid block. (5) Histological examination showed obliteration or narrowing of lumen of the small pial vessels involved in the adhesive arachnoiditis. In the cord parenchyma adjacent to the arachnoiditis, multiple spots of demyelination due secondary to ischemia demonstrated by LFB stain were noted. On the other hand, in the cord with the pia-arachnoid remained uninvolved, no demyelination was observed. (6) Localized adhesive arachnoiditis consisted of proliferation of fibrous tissue, lymphocytic infiltration and obliterating processes of small pial vessels involved in it. These data suggest that the cavitation within the cord would be induced by the ischemia, and hydromyelia would be produced by the pressure dissociation between the spinal subarachnoid space and the central canal.


[Surgical outcome of 32 cases in traumatic subdural hygroma].

32 cases of traumatic subdural hygroma (TSH) in adults with surgical treatment were retrospectively investigated by means by clinical features and CT findings. The cases consisted of 29 males and 3 females, aged 41 to 87 years (mean 69). Preparative CT scan of all cases revealed low density area and crescent shape in frontotemporal or frontoparietal subdural space. Half of the cases had bilateral lesions. At operation of TSH, color of subdural fluid collections was more water clear or xanthochromic than bloody. As a results, 22 out of 32 cases (69%) in TSH improved with surgical treatment. Many of effective cases of surgical treatment in TSH had short interval from trauma to operation and light disturbance of consciousness before operation. However, the other intracranial damage will also affect the clinical outcome of TSH, because the majority of cases in this study was accompanied by an intracranial damage including cerebral contusion, subarachnoid hemorrhage or intracranial hemorrhage. Nevertheless, surgical management for TSH was so effective that the operation should be undergone sooner interval from trauma, simultaneously considering the another intracranial lesions except TSH. But then, we experienced 7 cases (22%) of ventricular dilatation and 5 cases (16%) of chronic subdural hematoma in postoperative follow up CT scans. In 5 cases among the former, ventriculoperitonial shunt was done, and in 3 cases among the latter, burr hole evacuation was performed. Therefore, the postoperative course of TSH should require careful observation by CT scan and so on.


[A case of aneurysm of the peripheral middle cerebral artery presenting putaminal hemorrhage--pitfall in diagnosis].

A case mimicking hypertensive putaminal hemorrhage which was first treated by CT-guided stereotactic aspiration and eventually diagnosed as a sequence of the ruptured aneurysm of the left peripheral middle cerebral artery on postoperative angiography is presented. This 41 old, right-handed, hypertensive female suffered sudden onset of headache and right hemiparesis. Next day hematoma was evacuated by the CT-guided stereotactic aspiration because of progressive deterioration of the consciousness and patient recovered well. Fourteen days after the evacuation when she presented mild right hand weakness and motor aphasia, angiography revealed a saccular aneurysm with broad neck derived from left posterior parietal artery. A neck clipping necessitated an additional anastomosis between superficial temporal artery and distal middle cerebral artery because of parent artery stenosis. Postoperatively she is doing well with slight motor aphasia. Among the typical hypertensive putaminal hemorrhage diagnosed on CT scan, it is stressed that there may be a possibility of ruptured aneurysm situated on the peripheral middle cerebral artery.


Novel structural patterns in divalent cation-depleted surface layers of Aeromonas salmonicida.

The fish pathogen Aeromonas salmonicida possesses a regular surface layer (or A-layer) which is an important virulence determinant. The A-protein, a single bilobed protein organized in a p4 lattice of M4C4 arrangement with two morphological domains, comprises this layer. The role of divalent cations in the A-layer structure was studied to better understand A-protein subunit interactions affecting structural flexibility and function. Divalent cation bridges were found to be involved in the integrity of the A-layer. Two novel A-layer patterns were formed as the result of growth under calcium limitation or by chelation of divalent cations with EDTA or EGTA, thereby constituting the first reported case of formation of distinct regular arrays upon divalent cation depletion. Furthermore, under these conditions A-protein was sometimes released as tetrameric units, rather than in monomeric form. The formation of the two novel patterns is best explained by a sequence of structural rearrangements, following disruption of only one of the two A-layer morphological units, that is, those held together by divalent cation bridges. The free tetrameric units represent four A-protein subunits clustered around the unaffected four-fold axis.


Ultrastructural evidence for a triple structure of the lateral element of the synaptonemal complex.

This study describes composition and localization of several substructures of the synaptonemal complex (SC) using different techniques. The techniques which were used were surface spreading, critical point drying of isolated SCs, and sectioning of Lowicryl embedded testis material. The lateral elements (LEs) of the SC appear to be composed of three lateral substructures: two morphologically identical major strands and a third strand which is considerably thinner. The thinner strand is localized on the inner side of the two major strands of the lateral element. In late pachytene/early diplotene stages when the SC starts to disintegrate more than three strands can be observed in the LEs. A model is presented and the function of the different substructures is speculated upon.


Biochemical and ultrastructural studies of the C-type lectin bovine conglutinin.

The aim of this study was to correlate the supramolecular organization of conglutinin (BK) with its primary and tertiary structure and to gain more knowledge of functionally important regions of the molecule. BK analyzed by SDS-PAGE under standard reducing conditions (40 mM DTT) showed a major band at 43 kDa and weaker bands at 86 and 180 kDa. In contrast, reduction with 6-50 mM L-cysteine resulted in 37-kDa subunits indicating the presence of intrachain disulfide bonds within this subunit. Hydroxylamine treatment indicated presence of ester bonds in the 86- and 180-kDa subunits. Collagenase digestion and SDS-PAGE under reducing and nonreducing conditions resulted in bands of 20 and 15 kDa, respectively, indicating the presence of intrachain, rather than interchain, disulfide bonds in the carboxy terminus. Deglycosylation and glycan differentiation analysis of BK revealed the presence of O-linked glycans of GalNAc and alpha (2-3) linked sialic acid type, whereas no N-linked glycans were demonstrated. Binding experiments with GlcNAc-gold suggested that multivalency is required for carbohydrate binding to BK. Electron microscopy showed mostly tetramers, 96 nm in diameter, but also mono-, di-, and trimers were seen. The tetramers consisted of 40-nm strands, each with a peripheral globular head composed of subunits and connected to a common central lobe built from four ring-formed structures. The strands occasionally showed two bends, one close to the central lobe and another 25 nm from the lobe. These bends most likely correspond to the interrupted Gly-Xaa-Yaa repeats at residues 38 and 123.


MgATP binding changes neither the shape nor the flexibility of the heads of single myosin molecules.

The structural mechanism by which myosin heads exert force is unknown. One possibility is that the tight binding of the heads to actin drives them into a force-generating configuration. Another possibility is that the force-generating conformational change is inherent to the myosin heads. In this case the heads would make force by changing their shape according to the species of nucleotide in their active sites, the tight attachment to actin serving only to provide traction. To test this latter possibility, we used negative stain electron microscopy to search for a MgATP-induced shape change in the heads of single myosin molecules. We compared the heads of 10S smooth muscle myosin monomers (wherein MgATP is trapped at the active site) with the MgATP-free heads of 6S monomers. We found that to a resolution of about 2 nm, MgATP binding to the unrestrained myosin head does not drive it to change its shape or its flexibility. This result suggests that the head makes force by virtue of an induced fit to actin.


Polarity of microtubular structures in manchette-like formations: possible role of the "11th filament".

Bundles of microtubular structures appear in the cytoplasm of spermatids of the African frog Dicroglossus occipitalis. They are observed in the vicinity of axonemes. Natural tubulin polymerization leads to the formation of hooks on microtubular structures. They can be related to experimentally induced tubulin hooks. The direction of curvature of the hooks allows us to define the polarity of the bundles. This is opposite to the polarity of axonemal microtubules: Bundles and axonemes are antiparallel. Under colchicine action, arch-like microtubular structures are shown to open in the same direction as they lock. This enables us to characterize their opening and locking site: It corresponds to the place of the "11th filament" described in microtubular structures such axonemes. The "11th filament" is thus demonstrated to be the most susceptible to natural opening and to the action of colchicine in microtubular structures.


Decreased uptake of low density lipoprotein by LLC-PK cells cultured at low magnesium concentration.

LLC-PK cells grown in tissue culture on medium containing normal or deficient amounts of magnesium were presented with 125I-low density lipoprotein (LDL) and the uptake determined. Total LDL uptake by Mg-deficient cells decreased as the magnesium concentration decreased. The receptor binding of LDL and the internalization of LDL by the Mg-deficient cells were reduced. Degradation of LDL by Mg-deficient cells, when corrected for the reduced uptake, was less affected.


Assembly of 2-D membrane protein crystals: dynamics, crystal order, and fidelity of structure analysis by electron microscopy.

Membrane protein reconstitution into two-dimensional (2-D) ordered arrays is described. The assembly of 2-D crystals may be modeled as a two-step process: the membrane protein is first integrated in the lipid bilayer and then crystallized by removal of excess detergent or lipid and/or by precipitating agents. Lipid-detergent, protein-detergent, and lipid-protein interactions are critical during the first step, while lipid-protein and protein-protein interactions dominate events in the second step. The evidence supporting this model results from quasielastic light scattering analyses and electron microscopy of different lipid-detergent systems and reconstitution experiments with Escherichia coli porin OmpF, Phormidium laminosum photosystem I reaction centers, and integral membrane proteins of mammalian lens fiber cells.


Influence of endothelium in the in vitro vasorelaxant effect of magnesium on aortic basal tension in DOCA-salt hypertensive rat.

The objective of this study was to examine the influence of the endothelium on the extracellular magnesium induced relaxation of basal tension in isolated aortas from both mineralocorticoid-salt (DOCA-salt) hypertensive and control normotensive Sprague Dawley male rats. After incubation in magnesium-free physiological salt solution (PSS) (O mM magnesium), the increase of extracellular magnesium (1.2; 4.8 mM magnesium) caused a decrease in aortic tone which was significantly greater when endothelium was disrupted. Magnesium-induced relaxation was also more pronounced when endothelial NO production was blocked by 10(-4) M N omega-nitro-L arginine methyl ester (L-NAME). It is suggested that the vasorelaxation induced by extracellular magnesium is linked to the level of aortic basal tension developed in magnesium-free PSS. The endothelium does not seem to be directly implicated in magnesium-induced vasorelaxation in aortas from normotensive rats. However, in DOCA-salt hypertensive rats, the magnesium-induced relaxation of basal tension was less in the intact aorta (though not when the endothelium was disrupted) when the cyclo-oxygenase pathway was blocked by 10(-6) M indomethacin. These data therefore suggest that extracellular magnesium can promote relaxation by endothelium-dependent and cyclo-oxygenase-dependent mechanisms such as the production of relaxing prostacyclin in isolated aorta from DOCA-salt hypertensive rats.