Demonstration of Epstein-Barr viral DNA in paraffin-embedded specimens of lymphoproliferative syndrome. Evidence for a productive infection comparable to lymphoblastoid cell lines. B-cell lymphoproliferative syndromes (LPS) occurring in immunodeficient subjects are frequently associated with EBV infections. Histology as well as EBV-related serology are not diagnostic, but demonstration of EBV DNA in LPS suspected lesions might be useful for diagnosis. We studied four autopsied cases of LPS that developed in the setting of bone marrow transplantation, with proliferations ranging from poly- to monoclonal. Our protocol of DNA extraction allowed detection of EBV DNA in formalin-fixed, paraffin-embedded tissue specimens of all four cases. In dot blot hybridization the sensitivity in these specimens was 10% as compared to fresh frozen material, but still sufficient for a biotinylated probe. Southern blotting with the former DNA was not successful due to extensive degradation. In situ hybridization resulted in positive signals in all cases, using either 35S or 3H labeled probes. The labeling pattern suggested virus replication in B cells of LPS. By this, LPS resembles productively infected lymphoblastoid cell lines rather than latently EBV-infected Burkitt's lymphoma (BL). These findings strengthen the concept of LPS as a distinct clinicopathologic entity, differing from monoclonal, latently EBV-infected BL, as well as from polyclonal infectious mononucleosis, and the more common EBV-negative Non-Hodgkin lymphomas of the immunocompetent host.


Epithelial and mixed hepatoblastoma in the adult. Histological observations and general considerations. Two instances of hepatoblastoma in adults are reported, with one case representing a purely epithelial, the other a mixed epithelio-mesenchymal variant. The purely epithelial tumour, consisting entirely of so-called fetal elements, was present in the liver of a 35-year-old woman without any other hepatic changes, whereas the mixed tumour developed in the coarsely nodular liver of a 73-year-old man with a currently inactive cirrhosis. Besides the epithelial component, this case held predominantly indifferent spindle-shaped and histiocytic mesenchymal cells which frequently gave rise to osteoid and to bony trabeculae, and on occasion also to vascular structures, biliary ducts and even to groups of hepatocytes. The cytological and histological picture of both cases is comparable even in its details to that seen in the hepatoblastomas of early childhood. This concordance should be insisted upon as a diagnostic pre-condition if a mixed tumour in the adult, consisting of several components, is to be accepted as a hepatoblastoma. This similarity also enables us to recognize the purely epithelial variant of the hepatoblastoma in the older patient as belonging to this tumour category, and to separate it from other hepatic carcinomas. It is suggested that in these tumours the pluripotent cells, or the cells that have again become pluripotent, are derived from differentiated hepatocytes; no evidence in favour of the existence of a particular cellular subpopulation or of "stem cells" has been found.


Nonfunctioning pancreatic endocrine tumors. An immunohistochemical and electron microscopic analysis of 26 cases. Twenty-six patients with non-functioning pancreatic endocrine tumor (NFPET) were operated on during a 22-year period (1968-1990) at PUMC Hospital, Beijing. Of these, 19 were female and 7 were male with a mean age of 33 years. All these tumors, including 12 malignant and 14 benign, were solitary, and most of them were well-encapsulated. Immunohistochemical staining showed 23 (88.5%) containing 1-4 kinds of peptide hormone and 18 (69.2%) being multihormonal. In 7 of the 9 tumors subjected to electron microscopic study, various amounts of neurosecretory granules were found. Tumors of this series were clinically silent, but they contained some immunoreactive peptides, although the amount of the peptides varied from tumor to tumor.


Pleomorphic adenoma of the human female breast. We are presenting an interesting rare benign breast tumor which meets the characteristics of a salivary gland pleomorphic adenoma. The tumor was misdiagnosed during frozen section procedure, because several clusters, mainly composed of myoepithelial cells and surrounded by a chondroid matrix, were mistaken for cancerous blasts. Additionally the clinical and mammographic findings were very suspicious. Although this particular tumor is very infrequent, the pathologist should be aware of the difficulties in the differential diagnosis during frozen section and thus defer his final answer to the paraffin sections.


What's new in diagnosis and treatment of HPV-associated cervical lesions. The human papilloma virus (HPV) has been subject to intense discussion during the last decade, as evidence has accumulated to strongly suggest that viruses play an essential part in the genesis of cervical neoplasms. The contamination rate of the female population is reported to range between 5 and 80%, depending on the modality of detection. Many indications almost reliably suggest the oncogenicity of certain HPV-types. The highest progression rate was found in HPV 16 lesions, followed by HPV 18 lesions. In our material the incidence of HPV 16/18 and 31/33 infections compared to 6/11 was elevated in patients with cervical intraepithelial neoplasia (CIN) compared to women with non-CIN lesions. The exact histologic diagnosis of CIN lesions is essential for a differentiated therapy. Our data indicate that mild and moderate CIN lesions are detected more frequently by Pap smear and colposcopically directed punch biopsy than by cervical smears alone. Histological diagnosis and HPV-typing by in-situ hybridisation offers the possibility to establish a differentiated therapy, e.g. by way of close follow-up, local destruction or cone biopsy. Main attention should be directed at early detection of dysplasia, at timely institution of follow-up programs and a possibly conservative therapeutic management.


Rejection, herpesvirus infection, and Ki-67 expression in endomyocardial biopsy specimens from heart transplant recipients. We examined 164 endomyocardial biopsy specimens from 29 patients who received an orthotopic heart transplant since 1984. Rejection was graded according to the Hannover classification. Non-isotopic in situ DNA hybridization was conducted on cryostat sections with biotinylated probes for herpes simplex virus, Epstein-Barr virus, and cytomegalovirus. Serial sections of 126 biopsies were investigated immunohistochemically for the presence of activated T lymphocytes and proliferating cells, with monoclonal antibodies against interleukin 2 receptors (CD 25) and Ki-67 antigen. Relations between rejection grades, presence of proliferating cells, and presence of herpesvirus DNA were determined for the total number of biopsies. For some patients correlation of these parameters was studied over time. Herpesviral nucleic acids were detected in 25% of all biopsies: 37% of biopsies with relevant rejection (grades A 2 or A 3; 39% of all biopsies) compared to 18% of biopsies graded A 0, A 1, or A 5 (61% of all samples) (P less than 0.01). 56% of the biopsies with infection showed relevant rejection as compared with only 33% of the uninfected. Ki-67 expression was found in 41% of all biopsies, mainly in infiltrating cells: 69% of biopsies with relevant rejection compared with 23% of cases of minor/no rejection (P much less than 0.001). Ki-67 expression was also associated with herpesvirus infection: 66% of infected biopsies contained Ki-67 positive cells compared with 33% of uninfected biopsies (P less than 0.001). Herpesvirus infection was usually observed within the interstitial cell population, which, in many cases, displayed a considerable Ki-67 expression, too. In few cases only, hybridization was unequivocally found in vascular wall cells or myocytes. Viral myocarditis does not only mimic graft rejection morphologically, but it may also affect the course of rejection, via induction of antigenic changes or direct injury of cardiac tissues. Virus infections may also elicit or aggravate obliterative coronary artery disease, and thus contribute to accelerated graft atherosclerosis.


Human papillomavirus DNA in cervix. In-situ hybridization with biotinylated probes on Bouin's fixed paraffin embedded specimens. We examined retrospectively a series of 65 Bouin's fixed, paraffin-embedded tissue specimens from 8 condylomatous lesions, 16 condylomas associated with cervical intraepithelial neoplasia (CIN), and 12 neoplasia without condylomatous signs, for histological characteristics, the detection of viral structural antigen, the presence and typing of HPV DNA by molecular in situ hybridization with biotinylated probes types 6, 11, 16 and 18 under stringent conditions (Tm - 12 degrees C). HPV DNA was present in 34/65 (52%) specimens. Detection of viral structural antigen was positive in only 14% (3/22) specimens. HPV DNA were identified in 9/9 (100%) condylomatous lesions (with HPV type 6, 11, 18). Three condylomas were coinfected with both HPV type 6 or 11 and type 18; viral antigen was found in two specimens. HPV DNA were detected in 18/31 (58%) low grade and advanced CIN associated with condylomatous changes (type 6 = 5 specimens, type 11 = 3 specimens, type 16 = 4 specimens, type 18 = 6 specimens). Four of these cases were coinfected with both HPV type 6/11 and HPV type 16/18. Viral antigen was negative in all specimens. HPV DNA were detected in 7/25 (28%) advanced intra-cervical neoplasia (CIN III) without anatomopathological condylomatous changes (type 6 = 1 specimen, type 16 = 3 specimens, type 18 = 3 specimens). One of these specimens contained both HPV types 6 and 18. Viral antigen was found in one case. Our data confirm the association of HPV types 6 and 11 with condyloma and low grade neoplasia; HPV types 16 and 18 were associated with advanced cervical neoplasia.(ABSTRACT TRUNCATED AT 250 WORDS)


Simultaneous in situ hybridization for DNA and RNA reveals the presence of HPV in the majority of cervical cancer cells. Thirteen cases of invasive squamous cell carcinoma of the uterine cervix containing HPV types 16 or 18 DNA sequences, as detected by Southern blot analysis, were investigated by in situ hybridization on routine paraffin sections, using 35S nick-translated DNA probes. Simultaneous in situ hybridization for DNA and RNA showed that in ten out of 13 cases (77%) the percentage of tumor cells containing HPV 16 or 18 varied from 75 to 100%. In one case, harboring both in situ and invasive carcinoma, the same type of HPV DNA was detected in both components. This finding suggests that neoplastic cells retained the viral genome during progression to invasiveness.


Studies to determine whether an interaction exists among boron, calcium, and cholecalciferol on the skeletal development of broiler chickens. Two experiments were designed to determine the effect of dietary boron on broiler cockerels and four experiments were conducted to determine whether an interaction exists among dietary boron, cholecalciferol, and calcium. The parameters measured were weight gain, feed efficiency, tibia bone ash, rickets, tibial dyschondroplasia, and plasma minerals. All experiments were conducted with tibial dyschondroplasia-inducing basal diets fed to broiler cockerels from 1 to 16 days of age. Experiments 1 and 2 had four levels of dietary boron (0, 20, 40, and 80 mg/kg (Experiment 1) and 0, 5, 10, and 20 mg/kg (Experiment 2). Boron had no effect on weight gain, feed efficiency, or plasma minerals in either experiment. In Experiment 2, increasing levels of boron had no influence on tibial dyschondroplasia but did exert a quadratic effect on bone ash with 5 and 10 mg/kg boron increasing bone ash. In Experiment 1, bone ash and the incidence of tibial dyschondroplasia were unaffected, but the severity of tibial dyschondroplasia linearly increased by increasing boron levels. Experiments 3 to 6 had a 2 x 2 x 2 factorial arrangement of treatments with calcium at .65 and .90%, cholecalciferol at 110 and 1,100 ICU/kg, and boron at 0 and 40 mg/kg (Experiments 3 to 5) or 0 and 3 mg/kg (Experiment 6). The higher levels of calcium and cholecalciferol improved weight gain, decreased the incidence of rickets, and decreased the incidence and severity of tibial dyschondroplasia. Feeding cholecalciferol at 1,100 ICU/kg increased plasma calcium and plasma dialyzable phosphorus and decreased plasma magnesium. Calcium at .90% had no effect on plasma magnesium or plasma dialyzable phosphorus and increased plasma calcium only in Experiment 4. The only response to boron in Experiments 3 to 6 was a boron effect and a boron by cholecalciferol interaction on bone ash in Experiment 3, in which boron reduced bone ash at .65% calcium and 110 ICU/kg cholecalciferol. From these experiments, there is no indication that an interaction among boron, cholecalciferol, and calcium exists in broiler cockerels.


Stereoselective pharmacodynamic action of (+)-S-12967 and (-)-S-12968, isomers of a new slow acting 1,4-dihydropyridine derivative with calcium channel agonistic and antagonistic effects on rat aorta. The effects of (+)-S-12967 and (-)-S12968, isomers of a new dihydropyridine (1,4-DHP) derivative [2(7-amino 2,5-dioxaheptyl) 3-ethoxycarbonyl 4-(2,3-dichlorophenyl) 5-methoxycarbonyl 6-methyl 1,4-dihydropyridine] were studied on contractile responses of isolated thoracic aortas from rats and compared to that of nifedipine. The maximal relaxant effect of both isomers was reached in about 2 hr whereas the maximal relaxant effect to nifedipine was obtained within 30 min. The two 1,4-DPH isomers and nifedipine had a far more potent inhibitory effect on potassium (K+) than on noradrenaline (NA) induced contractions. They shifted the K+, Ca2+ and NA-concentration response-curves to the right and depressed the maximal vessel response to these agonists. Nifedipine was about 10 times more potent than the (-)-isomer which again was about 100 times more potent that the (+)-isomer. In contrast to nifedipine (-)-S-12968 and (+)-S-12967 had a dual action on K+ and Ca(2+)-induced contractions as both isomers in low concentrations, 3 x 10(-9)M and 3 x 10(-7)M, respectively, shifted the K(+)-concentration response curves to the left and increased the maximal response. In K(+)-depolarized preparations they increased the response to low Ca(2+)-concentrations without affecting the maximal vessel response at the highest Ca(2+)-concentrations. The result indicates that (+)-S-12967 and (-)-S-12968 possess Ca(2+)-agonistic as well as Ca-antagonistic properties. Compared to nifedipine both isomers are slow acting vasodilators. Their action as regards their potency is stereoselective and the (-)-isomer is more potent than the (+)-isomer.


Evidence for distinct phencyclidine and SKF10047 receptors following detergent treatment of rat brain membranes. In order to compare characteristics of benzomorphan and phencyclidine receptors, binding of the ligands [3H]SKF10047 and [3H]phencyclidine (PCP) was measured in intact rat synaptosomal membranes and in membranes treated with a detergent (CHAPS, a twitterionic derivative of cholic acid). Ligand binding was quantified in the particle containing fractions and in particle-free supernatants. About 20% of the SKF binding sites could be solubilized from the membranes by CHAPS under the conditions used here, while all PCP binding sites remained associated to particles. This observation and the inhibition patterns found for the two ligands indicate that the PCP receptors and the SKF receptors as delineated in this paper are indeed separate.


A purified recombinant organophosphorus acid anhydrase protects mice against soman. Since pharmacologic treatments of organophosphorus anticholinesterases (OPs) are nearing their practical limit other types of treatment are being sought. One approach is the prophylactic administration of scavengers that will detoxify OPs before they reach their critical target site. Using mice that were sensitized to OPs by depletion of their serum carboxylesterase with cresylbenzodioxaphosphorin oxide (CBDP), we have shown that animals pretreated intravenously with a purified organophosphorus acid anhydride hydrolase (parathionase) (0.10 mg per g body wt.) are not measureably affected by up to 34.4 micrograms soman per kg, a dose more than double that which is lethal to untreated animals. This result indicates that this approach is worthy of exploration and development for protecting military personnel and agricultural workers against OP intoxication.


[Comparison of a stat method of analysis of microorganisms for the presence of site-specific restriction endonuclease activity]. Sensitivity of several express-methods used for detection of bacterial endonucleases was compared. The most sensitive method is that employing Triton X-100.


Definition of the catalytic site of cytochrome c oxidase: specific ligands of heme a and the heme a3-CuB center. The three-subunit aa3-type cytochrome c oxidase (EC 1.9.3.1) of Rhodobacter sphaeroides is structurally and functionally homologous to the more complex mitochondrial oxidase. The largest subunit, subunit I, is highly conserved and predicted to contain 12 transmembrane segments that provide all the ligands for three of the four metal centers: heme a, heme a3, and CuB. A variety of spectroscopic techniques identify these ligands as histidines. We have used site-directed mutagenesis to change all the conserved histidines within subunit I of cytochrome c oxidase from Rb. sphaeroides. Analysis of the membrane-bound and purified mutant proteins by optical absorption and resonance Raman spectroscopy indicates that His-102 and His-421 are the ligands of heme a, while His-284, His-333, His-334, and His-419 ligate the heme a3-CuB center. To satisfy this ligation assignment, helices II, VI, VII, and X, which contain these histidine residues, must be in close proximity. These data provide empirical evidence regarding the three-dimensional protein structure at the catalytic core of cytochrome c oxidase.


[Determination of restriction endonucleases in Streptomyces and Nocardia colonies]. A simple technique is proposed for the detection of restriction endonucleases in Streptomyces and Nocardia cells. The analysis was performed directly in the cells collected from colonies cultivated on Petri dishes with an inoculation loop. The cells were treated with lysozyme, EDTA and Triton X-100. The lysates were tested for restriction endonucleases. The technique enables the detection of enzymes Nco I, Not I, Nru I, Sfr 3031, and Sfi I in the lysates of the respective strains-producers.


Zinc finger point mutations within the WT1 gene in Wilms tumor patients. A proposed Wilms tumor gene, WT1, which encodes a zinc finger protein, has previously been isolated from human chromosome 11p13. Chemical mismatch cleavage analysis was used to identify point mutations in the zinc finger region of this gene in a series of 32 Wilms tumors. Two exonic single base changes were detected. In zinc finger 3 of a bilateral Wilms tumor patient, a constitutional de novo C----T base change was found changing an arginine to a stop codon. One tumor from this patient showed allele loss leading to 11p hemizygosity of the abnormal allele. In zinc finger 2 of a sporadic Wilms tumor patient, a C----T base change resulted in an arginine to cysteine amino acid change. To our knowledge, a WT1 gene missense mutation has not been detected previously in a Wilms tumor. By comparison with a recent NMR and x-ray crystallographic analysis of an analogous zinc finger gene, early growth response gene 1 (EGR1), this amino acid change in WT1 occurs at a residue predicted to be critical for DNA binding capacity and site specificity. The detection of one nonsense point mutation and one missense WT1 gene point mutation adds to the accumulating evidence implicating this gene in a proportion of Wilms tumor patients.


Mechanism of dioxin action: Ah receptor-mediated increase in promoter accessibility in vivo. We have analyzed dioxin-inducible, Ah receptor-dependent changes in protein-DNA interactions at the CYP1A1 transcriptional promoter in intact mouse hepatoma cells. Our findings indicate that in uninduced cells, the promoter is inaccessible to its cognate binding proteins, which are known to be expressed constitutively. Dioxin induces, in Ah receptor-dependent fashion, an increase in promoter accessibility, which occurs rapidly and does not require ongoing transcription of the CYP1A1 gene. The change in promoter accessibility is not due to an altered pattern of cytosine methylation at the promoter; it probably reflects a 2,3,7,8-tetrachlorodibenzo-p-dioxin- induced change in the chromatin structure. These findings provide new insight into the mechanism of dioxin action and contribute to a better understanding of the regulation of inducible gene transcription in mammalian cells.


[Acid-base and ion-exchange properties of dietary fibers]. Acidic-basic characteristics of ionogenic groups of dietary fibers (DF) isolated from various raw materials were determined by potentiometric titration of DF using a method of individual weights and the ion-exchange properties of DF were estimated. The cation-exchanging ability of DF is within the range 0.2-1.5 meq/g DF. The calculated apparent constants of the acidic-basic dissociation of the DF functional groups are within the range allowing their reference to weakly acidic and basic groups. The source of the former can be polysaccharides (pectin substances, hemicelluloses), lignin and the protein accompanying DF components, which being an ampholyte possesses also basic properties. The effect of DF preparation on acidity of the gastric juice was studied to elucidate a possibility of using concentrated DF preparations for the correction of the states resulted from the hypersecretion of hydrochloric acid by the mucous membrane of stomach.


Expression pattern of the RAR alpha-PML fusion gene in acute promyelocytic leukemia. Two chimeric genes, PML-RAR alpha and RAR alpha-PML, are formed as a consequence of the acute promyelocytic leukemia (APL)-specific reciprocal translocation of chromosomes 15 and 17 [t(15;17)]. PML-RAR alpha is expressed as a fusion protein. We investigated the organization and expression pattern of the RAR alpha-PML gene in a series of APL patients representative of the molecular heterogeneity of the t(15;17) and found (i) two types of RAR alpha-PML mRNA junctions (RAR alpha exon 2/PML exon 4 or RAR alpha exon 2/PML exon 7) that maintain the RAR alpha and PML longest open reading frames aligned and are the result of chromosome 15 breaking at two different sites; and (ii) 10 different RAR alpha-PML fusion transcripts that differ for the assembly of their PML coding exons. A RAR alpha-PML transcript was present in most, but not all, APL patients.


Tumor necrosis factor soluble receptors circulate during experimental and clinical inflammation and can protect against excessive tumor necrosis factor alpha in vitro and in vivo. Tumor necrosis factor alpha (TNF alpha), a primary mediator of systemic responses to sepsis and infection, can be injurious to the organism when present in excessive quantities. Here we report that two types of naturally occurring soluble TNF receptors (sTNFR-I and sTNFR-II) circulate in human experimental endotoxemia and in critically ill patients and demonstrate that they neutralize TNF alpha-induced cytotoxicity and immunoreactivity in vitro. Utilizing immunoassays that discriminate between total sTNFR-I and sTNFR-I not bound to TNF alpha, we show that sTNFR-I-TNF alpha complexes may circulate even in the absence of detectable free TNF alpha. To investigate the therapeutic possibilities of sTNFR-I, recombinant protein was administered to nonhuman primates with lethal bacteremia and found to attenuate hemodynamic collapse and cytokine induction. We conclude that soluble receptors for TNF alpha are inducible in inflammation and circulate at levels sufficient to block the in vitro cytotoxicity associated with TNF alpha levels observed in nonlethal infection. Administration of sTNFR-I can prevent the adverse pathologic sequelae caused by the exaggerated TNF alpha production observed in lethal sepsis.


Possible involvement of the long terminal repeat of transposable element 17.6 in regulating expression of an insecticide resistance-associated P450 gene in Drosophila. P450-A and P450-B are electrophoretically defined subsets of cytochrome P450 enzymes in Drosophila melanogaster. P450-A is present among all strains tested, whereas expression of P450-B is associated with resistance to insecticides. Monoclonal antibodies were used to obtain cDNA clones for an enzyme from each P450 subset (i.e., P450-A1 and P450-B1). The P450-B1 cDNA was sequenced and shown to code for a P450 of 507 amino acids. Its gene has been named CYP6A2. Comparative molecular analyses of a pair of susceptible, 91-C, and resistant, 91-R, Drosophila strains were made. There was 20-30 times more P450-B1 mRNA in 91-R than in 91-C, and the small amount of P450-B1 mRNA in 91-C was significantly larger in size than that in 91-R. The P450-B1 gene in 91-R was structurally different from that in 91-C but was not amplified. The P450-B1 gene in 91-C contained a solitary long terminal repeat of transposable element 17.6 in its 3' untranslated region. It was absent in the P450-B1 gene of 91-R. On the basis of features of the long terminal repeat and its location in the gene of the susceptible fly, we propose that a posttranscriptional mechanism involving mRNA stability could be involved in regulating P450-B1 gene expression.


Molecular cloning of an atypical voltage-gated sodium channel expressed in human heart and uterus: evidence for a distinct gene family. Previously cloned voltage-dependent sodium channels exhibit a high degree of homology to one another and appear to comprise a single multigene family. We have now isolated and characterized cDNAs from both human adult heart and fetal skeletal muscle that encode a sodium channel alpha subunit that exhibits only moderate primary structure identity with other sodium channels and is prominently expressed in both heart and uterus. The approximately 7.2-kilobase cDNA sequence, designated hNav2.1, predicts a 1682-amino acid protein that bears 52%, 49%, and 46% overall identity with sodium channels cloned from rat brain, skeletal muscle, and heart, respectively. Positively charged S4 segments are present in hNav2.1, but there are fewer basic residues in repeat domains 1, 3, and 4 than in other cloned sodium channels. The cloning of hNav2.1 provides evidence for greater evolutionary divergence among voltage-dependent sodium channels and suggests that other sodium channel gene subfamilies may exist. The unique amino acid sequences in regions known to be involved in voltage-dependent activation and inactivation suggest that hNav2.1 will have novel gating properties.


Sequence analysis of the 5' noncoding region of hepatitis C virus. We have determined the nucleotide sequence of the 5' noncoding (NC) region of the hepatitis C virus (HCV) genome in 44 isolates from around the world. We have identified several HCV isolates with significantly greater sequence heterogeneity than reported previously within the 5' NC region. The most distantly related isolates were only 90.1% identical. Nucleotide insertions were seen in three isolates. Analysis of the nucleotide sequence from 44 HCV isolates in this study combined with that of 37 isolates reported in the literature reveals that the 5' NC region of HCV consists of highly conserved domains interspersed with variable domains. The consensus sequence was identical to the prototype HCV sequence. Nucleotide variations were found in 45 (16%) of the 282 nucleotide positions analyzed and were primarily located in three domains of significant heterogeneity (positions -239 to -222, -167 to -118, and -100 to -72). Conversely, there were three highly conserved domains consisting of 18, 22, and 63 completely invariant nucleotides (positions -263 to -246, -199 to -178, and -65 to -3, respectively). Two nucleotide domains within the 5' NC region, conserved among all HCV isolates studied to date, shared statistically significant similarity with pestivirus 5' NC sequences, providing further evidence for a close evolutionary relationship between these two groups of viruses. Additional analysis revealed the presence of short open reading frames in all HCV isolates. Our sequence analysis of the 5' NC region of the HCV genome provides additional information about conserved elements within this region and suggests a possible functional role for the region in viral replication or gene expression. These data also have implications for selection of optimal primer sequences for the detection of HCV RNA by the PCR assay.


An ATP-stabilized inhibitor of the proteasome is a component of the 1500-kDa ubiquitin conjugate-degrading complex. Proteins conjugated to ubiquitin are degraded by a 26S (1500-kDa) proteolytic complex that, in reticulocyte extracts, can be formed by the association of three factors: CF-1, CF-2, and CF-3. One of these factors, CF-3, has been shown to be the proteasome, a 650-kDa multicatalytic protease complex. We have purified a 250-kDa inhibitor of the proteasome and shown that it corresponds to CF-2. In the presence or absence of ATP, this factor inhibited hydrolysis by the proteasome of both fluorogenic tetrapeptides and protein substrates. When the inhibitor, proteasome, and CF-1 were incubated together in the presence of ATP and Mg2+, degradation of ubiquitin-125I-lysozyme occurred. Both the inhibitory activity and the ability to reconstitute ubiquitin-125I-lysozyme degradation were very labile at 42 degrees C, but both activities were stabilized by ATP or a nonhydrolyzable ATP analog. SDS/PAGE indicated that the 250-kDa inhibitor fraction contained a major subunit of 40 kDa (plus some minor bands). The 125I-labeled inhibitor and purified proteasome formed a complex. When CF-1, ATP, and Mg2+ were also present, the 125I-labeled inhibitor along with the proteasome formed a complex of 1500 kDa. The inhibitor (CF-2) thus appears to be an ATP-binding component that regulates proteolysis within the 1500-kDa complex.


Molecular cloning of the alpha-1 subunit of an omega-conotoxin-sensitive calcium channel. Of the four major types of Ca channel described in vertebrate cells (designated T, L, N, and P), N-type Ca channels are unique in that they are found specifically in neurons, have been correlated with control of neurotransmitter release, and are blocked by omega-conotoxin, a neuropeptide toxin isolated from the marine snail Conus geographus. A set of overlapping cDNA clones were isolated and found to encode a Ca channel alpha-1 subunit, designated rbB-I. Polyclonal antiserum generated against a peptide from the rbB-I sequence selectively immunoprecipitates high-affinity 125I-labeled omega-conotoxin-binding sites from labeled rat forebrain membranes. PCR analysis shows that, like N-type Ca channels, expression of rbB-I is limited to the nervous system and neuronally derived cell lines. This brain Ca channel may mediate the omega-conotoxin-sensitive Ca influx required for neurotransmitter release at many synapses.


Pregnenolone biosynthesis in C6-2B glioma cell mitochondria: regulation by a mitochondrial diazepam binding inhibitor receptor. The C6-2B glioma cell line, rich in mitochondrial receptors that bind with high affinity to benzodiazepines, imidazopyridines, and isoquinolinecarboxamides (previously called peripheral-type benzodiazepine receptors), was investigated as a model to study the significance of the polypeptide diazepam binding inhibitor (DBI) and the putative DBI processing products on mitochondrial receptor-regulated steroidogenesis. DBI and its naturally occurring fragments have been found to be present in high concentrations in C6-2B glioma cells, to compete against specific isoquinolinecarboxamide or 4'-chlorodiazepam binding to mitochondrial recognition sites with high affinity, and to stimulate mitochondrial pregnenolone formation. These data suggest that this cell type may express both the receptor and the putative agonist ligand to regulate steroidogenesis. Therefore, we propose to term this mitochondrial receptor MDR (mitochondrial DBI receptor) to indicate its responsiveness to DBI in steroid biosynthesis. In the present work, we show that mitochondria of C6-2B cells convert (22R)-22-hydroxycholesterol to pregnenolone by a mechanism blocked by aminoglutethimide. Immunoblotting confirmed the presence of relatively high levels of cytochrome P-450 cholesterol side-chain-cleavage enzyme in C6-2B cell mitochondria. Furthermore, isoquinolinecarboxamide binding sites associated with the 18-kDa mitochondrial polypeptide subunit of the MDR are abundant in C6-2B glioma cell mitochondria (Bmax approximately 30 pmol/mg protein) and are coupled to the regulation of steroid biosynthesis. Occupancy of MDRs with nanomolar concentrations of the naturally occurring polypeptide, DBI, as well as its naturally occurring processing product tetratriacontaneuropeptide [DBI-(17-50)] increases pregnenolone formation. Clonazepam and octadecaneuropeptide [DBI-(33-50)], which exhibit a higher affinity for gamma-aminobutyric acid type A receptors but a low affinity for MDR, were ineffective in stimulating pregnenolone synthesis. These findings provide evidence that C6-2B cells exhibit a significant steroidogenic activity which resembles that found in peripheral endocrine organs and they suggest that MDRs and DBI are involved in the regulation of glial cell steroidogenesis.


Regulation of pregnenolone synthesis in C6-2B glioma cells by 4'-chlorodiazepam. An experimental model to study synthesis of cholesterol and pregnenolone from the precursor mevalonolactone (MVA) was developed in C6-2B glioma cells. The steroidogenic capability of this cell line and the regulation of pregnenolone production by 4'-chlorodiazepam (4'CD), a specific ligand for the mitochondrial diazepam binding inhibitor (DBI) receptor (MDR), were investigated. Cells maintained in serum-free media were incubated with lovastatin (20 microM) and two inhibitors of pregnenolone metabolism, trilostane (25 microM) and 1,2,3,4-tetrahydro-4-oxo-7-chloro-2-naphthylpyridine (10 microM). Under these conditions the incorporation of [3H]MVA into cholesterol and pregnenolone formation was biphasic, with an initial rapid phase (within 1 min) followed by a slower phase. Cholesterol and pregnenolone were identified by coelution with authentic steroids from a Si 60 Lichrosorb column and gas chromatography/mass spectrometry. Pregnenolone synthesis in intact C6-2B glioma cells was stimulated by nanomolar concentrations of 4'CD after 5 min of incubation with MVA. The stimulatory effect was dependent on drug concentration and the maximal effect was achieved at 10 nM. The time course showed that the incorporation of MVA into pregnenolone is accelerated by the MDR ligand. Cholesterol synthesis is only slightly and not significantly affected by 4'CD. These results support the view that steroid synthesis occurs in a glioma cell line. Moreover, we provide evidence for a rapid steroid synthesis in C6-2B glioma cells, which in turn appears to be accelerated by 1-100 nM 4'CD, a MDR ligand.


Divergent viral superantigens delete V beta 5+ T lymphocytes. Several murine superantigens in association with class II major histocompatibility complex proteins have been shown to cause the deletion of T cells based on the expression of particular beta-chain variable region (V beta) gene segments. We have previously shown that mice expressing the Etc-1 superantigen, encoded by an open reading frame within the 3' long terminal repeat of the endogenous mouse mammary tumor provirus (Mtv), Mtv-9, delete T cells expressing either V beta 5 or V beta 11 gene segments. Comparison of several Mtv 3' long terminal repeat open reading frame sequences has indicated that the carboxyl terminus likely encodes the V beta specificity of these proteins. Our analysis of C57BL/6 x DBA/2 recombinant inbred strains of mouse revealed three Mtv-9-negative strains that nevertheless have a low frequency of V beta 5-expressing T cells. Here we demonstrate that a second endogenous superantigen, responsible for the deletion of V beta 5-bearing T cells, is encoded by a gene mapping to Mtv-6 on chromosome 16. Surprisingly, the carboxyl-terminal sequences of the Mtv-6 and -9 superantigens are extremely divergent, in spite of the fact that they both mediate the deletion of V beta 5+ lymphocytes.


Conditioned place preference induced by Ro 16-6028, a benzodiazepine receptor partial agonist. A place conditioning situation was used to assess the putative affective properties of benzodiazepine receptor ligands in the rat. The benzodiazepine receptor partial agonist Ro 16-6028 induced a conditioned place preference, suggesting that this compound has rewarding properties. The benzodiazepine receptor antagonist Ro 15-1788 induced neither place preference nor aversion, but prevented the place preference induced by Ro 16-6028, suggesting that the rewarding effects of Ro 16-6028 are due to its action on the benzodiazepine receptor. The benzodiazepine receptor full agonist diazepam did not induce a conditioned place preference in our hands, in contrast with previous studies. The sensitivity of place conditioning with benzodiazepine ligands to situational factors, such as the existence of a preconditioning preference, is discussed.


Solvent dielectric effects on protein dynamics. Electron paramagnetic resonance (EPR) spectroscopy and molecular dynamics (MD) simulations were used to investigate the dynamics of alpha-chymotrypsin in solvents ranging in dielectric constant from 72 to 1.9. EPR measurements showed that motions in the vicinity of two spin-labeled amino acids (Met-192 and Ser-195) decreased dramatically with decreasing solvent dielectric constant, a trend consistent with changes in the electrostatic force between charged residues of the protein. EPR results and MD simulations revealed a very similar functional dependence between rates of motion in the protein and the dielectric constant of the bulk solvent; however, predicted motions of protein atoms were markedly faster than measured motions of the spin labels. MD calculations for dielectric constants of 5 and 72 showed the greatest differences near the outer surface of the protein. In general, at the lower dielectric constant many atoms of the protein move more slowly, and many of the slowest residues are near the exterior. These results suggest that altered dynamics may contribute to the unusual properties--e.g., modified stereoselectivities--of enzymes in nearly dry organic solvents.


Human and dog erythrocytes: relationship between cellular ATP levels, ATP consumption and potassium concentrations. The intracellular K+/Na+ ratio of various mammalian cell types are known to differ remarkably. Particularly noteworthy is the fact that erythrocytes of different mammalian species contain entirely different potassium and sodium concentrations. The human erythrocyte is an example of the supposedly "normal" high potassium cell, while the dog erythrocyte contains ten times more sodium than potassium ions (Table I). Furthermore, this difference is sustained despite the plasma sodium and potassium concentrations being almost identical in both species (high Na+ and low K+). In spite of these inorganic ion differences, both human and dog erythrocytes contain 33% dry material (mostly Hb) and 67% water. Conventional cell theory would couple cellular volume regulation with Na+ and K+ dependent ATPase activity which is believed to control intracellular Na+/K+ concentrations. Since the high Na+ and low K+ contents of dog erythrocytes are believed to be due to the lack of the postulated Na/K-ATPase enzyme, they must presumably have an alternative mechanism of volume regulation, otherwise current ideas of membrane ATPase activity coupled volume regulation need serious reconsideration. The object of our investigation was to explore the relationship between ATPase activity, ATP levels and the Na+/K+ concentrations in human and dog erythrocytes. Our results indicate that the intracellular ATP level in erythrocytes correspond with their K+, Na+ content. They are discussed in relation to conventional membrane transport theory and also to Ling's "association-induction hypothesis", the latter proving to be a more useful basis on which to interpret results.


Maturation of rat spermatozoa: ESR spin labeling studies. The ability of spermatozoa to reduce nitroxide spin--TEMPO has been used as a parameter to understand maturation, capacitation and calcium uptake of sperm obtained from Holstman strain rats. The rate of spin label reduction by sperm follows the trend--caput greater than cauda greater than corpus. With the increase in age, the electron donating capability shows first a gradual increase, for 60- to 85-day-old rats, peaking at 85 days (corresponding to puberty) and leveling off after 92 days. Calcium uptake takes place in two phases which corresponds to accumulation of and activation by calcium. The presence of polyclonal antibody which is known to cause agglutination, does not adversely affect the sperm activity.


Chronic intrahypothalamic infusions of insulin or insulin antibodies alter body weight and food intake in the rat. In Experiment 1, one-week infusion of insulin (0.15, 1.5, or 15.0 microU/hr) into the ventromedial hypothalamus (VMH) of rats reduced body weight (BW) and nighttime food intake (FI). While 0.15 microU/h decreased daytime FI, 1.5 microU/h increased daytime FI and 15.0 microU/h left daytime FI unchanged. Total daily FI was decreased by the two highest doses. In Experiment 2, intra-VMH infusion of specific insulin antibodies (1.5 microUeq/h) increased BW and FI, while C-peptide antibodies were ineffective. In Experiment 3a, intracerebroventricular infusions of insulin failed to decrease FI and BW comparably to similar intrahypothalamic infusions. In Experiment 3b, intra-VMH insulin was infused via cannulae that bypassed the cerebral ventricles. The decrease in FI and BW was comparable to that observed when insulin was infused via cannulae that penetrated a ventricle. Histology from animals used in Experiments 1-3 indicates that optimum sites for insulin-induced changes in BW and FI in the hypothalamus lie in an area that includes portions of the paraventricular, arcuate, dorsomedial, and ventromedial nuclei.


Sleep deprivation-induced hyperthermia following antigen challenge due to opioid but not interleukin-1 involvement. Eight hours total sleep deprivation does not affect colonic temperature. The combination of a subpyrogenic challenge of sheep red blood cells with sleep loss however, can produce a significant rise in colonic temperature that peaks during the third hour of the sleep deprivation vigil. The regulation of this increase in colonic temperature appears to be opioid in nature and not because of the release of the cytokine interleukin-1. It would appear that the combination of sleep loss and low dose antigen challenge, both manipulations of themselves nonpyrogenic, produces a synergistic rise in colonic temperature. The implication of a psychologically-derived stress response via the opioid system may explain this finding.


Naloxone-sensitive potentiation at granule cell synapses in the ventral dentate gyrus and stress ulcers. Continuous perfusion of the granule cells in the ventral dentate gyrus with naloxone blocked the long-term potentiation (LTP) induced by high-frequency electrical stimulation of the posterolateral amygdala in freely-moving rats. This treatment also aggravated the stomach ulceration produced by cold restraint. LTP, on the other hand, attenuated the gastric stress pathology. It was suggested that a naloxone-sensitive granule cell gate modulates the impact of environmental stressors.


Plasma testosterone concentration in adult male rats following acquisition of copulation-illness associations. Prior research has identified stimuli and procedures that elicit increments in plasma testosterone in copulating male rats. In the present experiment, we demonstrate that associative inhibition of copulatory behaviors in male rats is not correlated with and cannot be attributed to a conditioned suppression of testosterone. Each male rat was paired with an inaccessible estrous female for 7 min and was then given an opportunity to copulate. Two groups received an injection of either lithium chloride (LiCl; 0.3 M, 20 ml/kg, IP) or saline (0.3 M, 20 ml/kg, IP) immediately after each of 11 pairings spaced at 3- to 4-day intervals. A third group received a noncontingent injection of LiCl 24 h after each pairing. After an initial screening for copulatory behaviors, a fourth group received only handling comparable to that received by the other three groups. Rats that received contingent LiCl gradually ceased to copulate; rats that received either noncontingent LiCl or saline remained vigorous copulators. Male rats were returned to their home cages on Trial 12 after 7-min exposure to an inaccessible female. Blood was collected by decapitation 38 min later. Testosterone levels, measured by radioimmunoassay, were significantly higher for saline than for handled control rats. Testosterone levels for handled control rats, however, were comparable to those of copulating and noncopulating rats that had received either noncontingent or contingent LiCl, respectively.


Neuroendocrine evidence for decreased function of alpha 2-adrenergic receptor after electroconvulsive therapy. Growth hormone (GH) and hypotensive responses to clonidine (150 micrograms, i.v.) were investigated before and after electroconvulsive therapy (ECT) in 16 depressed patients. Because of high baseline serum GH concentrations, results from only 10 patients could be evaluated. The level of GH secretion induced by clonidine was significantly reduced after ECT, but the hypotensive responses to clonidine remained unchanged. The results indicate downward regulation of the sensitivity of alpha 2-adrenergic receptors in the hypothalamus after ECT.


Variation of RBE between p(75) + Be and d(50) + Be neutrons determined for chromosome aberrations in Allium cepa. The RBE of p(75) + Be neutrons relative to d(50) + Be neutrons has been determined for chromosome aberrations induced in Allium cepa (onion) roots. Two biological criteria were selected: the average number of aberrations (mainly fragments) per cell in anaphase and telophase, and the percentage of aberration-free cells. The influence of sampling time (3 to 7 h incubation) between irradiation and fixation was investigated systematically. This factor did not significantly influence the results. The RBE values of p(75) + Be neutrons compared to those of d(50) + Be neutrons were 0.85 (0.79-0.91) and 0.87 (0.80-0.95) for the first and the second criteria, respectively. In previous experiments for the same beams, we found an RBE of 0.90 (0.86-0.94) for survival of V79 cells (D0 ratio), 0.96 (0.93-0.99) for the intestinal crypt cell system, and 0.83 (0.70-0.96) for Vicia faba growth delay.


[Extended pulmonary resection in non-small cell lung cancer]. The authors give an account of their experience with extensive pulmonary resections on account of an extensive non-small-cell carcinoma of the lungs. During a 13-year period (1976-1988) they made 140 extensive resections of the lungs. As to histological characteristics, the epidermoid type of tumour predominated. The tumour spread most frequently to mediastinal nodes and the thoracic wall. The mean age of the patients was 54.9 years, the mortality within 30 days after operation 5.7%. The five-year survival of patients after operation was evaluated in patients operated between 1976 and 1988 when 90 operations were performed. The majority of patients (32.5%) survived combined surgery. Two patients survive after 10 years. Due to the small number of patients and their difficult comparability it is not possible to evaluate the effectiveness of different types of treatment statistically. In the literature views on combined postoperative treatment differ, a randomized study would be useful. In the conclusion the authors advocate continuation of the hitherto used trend of therapy, they recommend introduction of new chemotherapeutic agents in close collaboration with oncologists and concentration of patients in specialized departments.


[Malignant cystosarcoma phyllodes].

[Malignant cystosarcoma phyllodes]. The authors deal with the clinical and pathological aspects and histopathological criteria of malignity of cystosarcoma which are important from the aspect of assessment of the extent and type of surgical intervention. Large malignant cystosarcomas and cystosarcomas taking up the whole breast with damage of the skin are indicated for simple mastectomy, while small malignant cystosarcomas (i. e. in relation to the size of the breast) can be treated by a wide local excision with a safety margin, as some primary as well as recurrent phylloid tumours behave in a benign way or may transform into a fibroadenoma or a dysplastic focus. Moreover contrary to carcinoma of the breast, there is no association with a multicentric character of the disease. Axillary lymph nodes are only rarely affected; therefore dissection of the nodes is not necessary.


Multiple rearrangements and activated expression of c-myc induced by woodchuck hepatitis virus integration in a primary liver tumour. Woodchuck hepatitis virus (WHV) is a small, partially double-stranded DNA virus. Like the related human hepatitis B virus (HBV), WHV induces acute and chronic hepatitis and hepatocellular carcinoma (HCC) in its natural host. WHV DNA integration into c-myc and N-myc, resulting in deregulated expression of these genes, has been described previously in woodchuck HCC. We have analysed a woodchuck liver tumour in which WHV DNA was integrated in the c-myc gene. The virus insertion provoked multiple alterations in one c-myc allele, probably involving secondary deletions and mutations. Integrated viral DNA, including promotor and enhancer sequences, acted as an insertional mutagen, leading to enhanced expression of heterogenous c-myc transcripts ranging from 7.2 to 14 kb in size, strikingly longer than normal 2.3-kb c-myc RNA. These results provide an additional example in which the oncogenic activation of a myc gene by cis-acting effect of WHV insertion may play a critical role in virus-induced woodchuck HCC.


Intracranial metastasis from a spinal cord primitive neuroectodermal tumor: case report. A patient with intracranial seeding from a spinal cord primitive neuroectodermal tumor with ependymal differentiation is presented. The first and second stages of intracranial dissemination were well controlled by a combination of irradiation and chemotherapy. The authors review previously published cases and discuss the possible mechanism of seeding from the spinal cord to the intracranial region.


Corticotroph cell hyperplasia in a patient with Addison disease: case report. A woman with Addison disease developed hyperpigmentation, headache, and nausea despite conventional replacement therapy with cortisone. Excessively elevated plasma adrenocorticotropic hormone (ACTH) with absence of response to administration of corticotropin-releasing factor (CRF), and roentgenological evidence of enlargement of the sella turcica, as well as detection of enlarged pituitary gland on magnetic resonance images, led to a diagnosis of ACTH-producing microadenoma, which was removed by transsphenoidal microsurgery. The specimen obtained at surgery evidenced corticotroph hyperplasia, as demonstrated by immunohistochemical staining for ACTH. Fine structure exhibited densely granulated cells with a few bundles of microfilaments and an abundance of large lysosomal bodies. Surgical removal of the hyperplasia alleviated the patient's symptoms, and hyperpigmentation faded remarkably. Her plasma ACTH level returned to normal, has remained normal for more than 3 years, and responds adequately to CRF administration.


Primary intracerebral malignant lymphoma associated with different histological types of carcinoma: report of two cases. Two rare cases with histologically proven multiple primary neoplasms are described: an association of intracerebral malignant lymphoma with hepatocellular carcinoma in one case and with squamous cell carcinoma of the uterine cervix in the other. Therapeutic problems pertinent to the coexistence of primary intracerebral malignant lymphoma and neoplasms of a different histological type are discussed.


Surgical treatment of intrahepatic cholangiocarcinoma: four patients surviving more than five years. To find the rational surgical strategy for the treatment of intrahepatic cholangiocarcinoma (ICC), clinical features of ICC were studied in 20 patients who underwent hepatic resection in the National Cancer Center Hospital from 1980 to 1990. According to the morphologic pattern, we classified the ICCs into two subcategories, mass-forming and infiltrating, which correlated with their biologic behavior. Of 10 patients who underwent hepatectomy for mass-forming ICC, three survived more than 5 years without recurrence. The 1-, 3-, and 5-year survival rates were 59.3%, 44.4%, and 44.4%, respectively. Of 10 patients who underwent hepatectomy for infiltrating ICC, one survived more than 5 years without recurrence. The 1-, 3-, and 5-year survival rates were 72.0%, 27.0%, and 27.0%, respectively. The pathologic findings and recurrences indicated that the salient feature of the mass-forming type was its tendency for intrahepatic metastasis especially near a main lesion, and of the infiltrating type was the infiltrative spread via Glisson's capsule and hilar lymph nodal metastasis. An anatomic and extensive liver resection should be performed for mass-forming ICC, whereas a hepatectomy with excision of the extrahepatic bile duct and hilar lymph nodal dissection is recommended for infiltrating ICC.


An isolated caudate lobectomy by the transhepatic approach for hepatocellular carcinoma in cirrhotic liver. A new strategy for the treatment of hepatic malignancy located deep in the cirrhotic liver is reported. A 66-year-old man, who had a 3 cm hepatocellular carcinoma in the cranial part of the caudate lobe, underwent an isolated caudate lobectomy. By transecting along the interlobar plane and opening the hepatic hilus anteriorly, we resected almost the entire caudate lobe without loss of the remaining liver parenchyma.


Proofreading, NTPases and translation: successful increase in specificity. The discussion of proofreading started in the April issue of TIBS is completed by treating the two branched Michaelis-Menten enzymes that can proofread. The conditions required for proofreading can be seen to determine the expression of proofreading in its biological settings. There are surely instances of proofreading as yet unrecognized.


Lymphosarcoma in experimentally induced feline immunodeficiency virus infection [corrected]. A cat experimentally infected with feline immunodeficiency virus (FIV) but known to be free of feline leukaemia virus (FeLV) developed lymphosarcoma. The lesions in the liver and kidneys were present nine months after infection, when the cat was 21 months old. The cat had no overt signs of immunodeficiency and it is suggested that the B cell activation induced shortly after FIV infection produced a large pool of proliferating lymphocytes from which the malignant cells emerged.


Replication and immunosuppressive effects of Pseudorabies virus on swine peripheral blood mononuclear cells. The infectivity and potential immunosuppressive effects of Pseudorabies virus (PRV) was evaluated in swine peripheral blood mononuclear cells (PBMC). Virus progeny titers and viral DNA synthesis at various intervals post-inoculation revealed the replication of PRV in both peripheral blood monocytes and lymphocytes; however, replication in lymphocytes was restricted compared with monocytes. PRV infection resulted in the damage and death of monocytes. Although PRV did not appear to affect the viability of the lymphocytes, PRV infection suppressed lymphocyte functions such as proliferation and interleukin-2 (IL-2) synthesis in response to Concanavalin A. This immunosuppression was dependent upon the multiplicity of infection (MOI) of infectious PRV. UV-inactivated PRV was not immunosuppressive. There was no effect of PRV on natural killer (NK) cell activity. The reduction of lymphocyte proliferation by PRV was not reversible by the addition of supernatant containing porcine IL-2 and non-infected monocytes to the infected cultures. The results from these in vitro studies demonstrate that PRV can infect and cause immunosuppressive effects on swine PBMC. These effects may explain the potential role of PRV in predisposing infected pigs to secondary infection and support the hypothesis that PRV can spread systemically by infected PBMC in blood and lymph.


In vitro responses of cheetah mononuclear cells to feline herpesvirus-1 and Cryptococcus neoformans. In vitro T cell function by domestic cats and cheetahs to two common pathogens, feline herpesvirus-1 (FHV-1) and Cryptococcus neoformans, was assessed. Peripheral blood mononuclear cells (PBM) were stimulated with two strains of UV-inactivated FHV-1, whole heat-killed organisms or capsular antigen of Cryptococcus neoformans, and proliferative responses measured. As a group, cheetah PBM responded significantly poorer than domestic cat PBM when cultured with FHV-1. However, individual cheetah responses varied widely. Supplementation of cultures with exogenous interleukin 2 (IL-2) significantly increased the level of response of individual cheetahs to both strains of FHV-1. Cheetah sera contained slightly higher neutralizing antibody titers to FHV-1 than did domestic cat sera, suggesting that B cells function adequately in cheetahs. When stimulated with Cryptococcus neoformans, both species had similar incidences of positive proliferative responses. These data demonstrate that cheetahs exhibit heterogeneous responses to specific antigens, similar to domestic cats. However, a lower group response to FHV-1 in cheetahs suggests species differences occur. In addition, level of variability in major histocompatibility complex (MHC) class I-like genes, as determined by Southern blot hybridization, does not appear to correlate with a uniform response in in vitro functional assays. Therefore, additional mechanisms influence the final outcome of the immune response.


A histopathological and ultrastructural study of eccrine porocarcinoma with special reference to its subtypes. Five cases of eccrine porocarcinoma were studied by light and electron microscopy. Histopathologically, these could be classified into two types; the common and the giant cell type. The common type was characterized by almost uniform medium-sized cuboidal tumour cells and a formation of well-developed intracytoplasmic lumina. A broad diversity of histopathological and ultrastructural features was seen in these tumours. The tumours of the giant cell type consisted of mononuclear polygonal cells and bizarre giant cells. This type was considered to be an undifferentiated form of porocarcinoma.


Malignant rhabdoid tumour of the uterus: an immunohistochemical and ultrastructural study. Malignant rhabdoid tumours (MRTs) are highly aggressive neoplasms which most frequently occur in the kidney of young children. Several cases of primary MRT occurring in extra-renal sites have been reported, particularly in the soft tissues. We report a case of primary MRT of the uterus, a very rare site for this neoplasm, with morphological, immunohistochemical and ultrastructural features corresponding to restrictive morphological criteria for MRT. The possible differential diagnoses were considered.


[Determination of differential renal function using 99m Tc-DMSA scintigraphy]. By examination of 331 patients, using 99mTc-DMSA, the authors confirmed that static scintigraphy is an accurate method for assessment of the separate renal function. In four groups of patients with different positions of the kidneys the authors proved that correction for different absorption of gamma radiation from the left and right kidney is essential. Without correction results with a significant error are obtained, in particular in patients with an asymmetric position of the kidneys, where the results calculated without correction cannot be used for clinical purposes. The results are not influenced by the time interval between administration of the radiopharmaceutical preparation and examinations within an interval of 2 to 4 hours, with the exception of patients with severe obstructive uropathy. For assessment of the relative function of the kidneys it is better to examine the patient in a vertical position. The reproducibility of results in repeated examinations and repeated evaluation is excellent.


[Rational neurologic diagnosis in fecal incontinence]. The effective therapy of a disturbance of anal continence requires an adequate preoperative diagnostics. In most cases this includes some kind of neurophysiological investigation. Close cooperation between the internist, the surgeon and the neurologist is advantageous. The initial history, clinical examination and electromyography of the pelvic floor will usually be enough to differentiate between aetiologies. In the first phase it is necessary to decide whether the problem is neurogenic or muscular, and then to see localising signs for a nerve defect (central or peripheral) or, respectively, signs of a defect in the skeletal musculature. It must be said that a peripheral nerve lesion (eg. a stretching injury of branches of the pudendal nerve) and a muscular defect may be combined. At the end of the diagnostic process the surgeon or internist and neurologist must decide together whether the diagnosis is appropriate, and then whether an operative or non-operative approach to treatment is fitting. In the second phase of diagnosis are further neurophysiological investigations, which are only indicated in more special circumstances. These investigations include: nerve conduction velocities, reflex latencies (anal-, bulbocavernosus-, and pudendoanal reflexes), evoked potentials, and the single fibre EMG to determine fibre density. These neurophysiological investigation in proctology allows the clinician a wider scope of diagnostic possibilities, which should lead to more sensible therapeutic options being taken.


[Cervix cytology in females with kidney transplants]. Cervical dysplasia has been reported to occur more frequently in female renal transplant patients. In a 4 year surveillance-period no new cases of dysplasia or invasive cancer were found by cytological or histological methods. Despite the low incidence of abnormalities the use of cytological screening is recommended, above all in the view of a slow progress of the disease.


Age-dependent attenuation of the decrease of C fibers by capsaicin and its effects on responses to nociceptive stimuli. The effects of subcutaneous treatment of mice 10, 15, or 20 days (young) or 30 or 60 days (adult) of age with capsaicin on development of unmyelinated (C) fibers in the L4 dorsal roots were examined. The responses of the mice 2-4 months later to thermal (hot plate; 55 degrees C) and neurogenic plasma extravasative (chemogenic nociception) stimuli were evaluated. Capsaicin treatment 10 days after birth affected development of myelinated fibers significantly (7.9% reduction). Capsaicin treatment 10, 15, 20, and 30 days after birth reduced the number of C fibers 11.0-51.7% and even treatment 60 days after birth caused a significant decrease (10.0%) in the mean number of C fibers. The destruction of C fibers by capsaicin was attenuated during development, but individual differences in the reduction of C fibers were observed on and after 15 days of life and seemed to become more marked over time. Neurogenic plasma extravasation related to primary sensory neurons was decreased by capsaicin, irrespective of the time of treatment. In parallel with reduction of C fibers, hot-plate latency was increased significantly by treatment of young animals with capsaicin. These results suggest that the effect of capsaicin on thermal nociception is age-dependent and is correlated with decrease of C fibers. However, a marked increase in hot-plate latency did not always correspond to a marked decrease of C fibers. In contrast, the reduction of plasma extravasation was not age-dependent: Reduced extravasation of dye persisted during development.(ABSTRACT TRUNCATED AT 250 WORDS)


The pars tuberalis of the hypophysis: a modulator of the pars distalis? The pineal hormone melatonin is known to influence circadian systems. Melatonin is also ascribed to mediate photoperiodic effects on the regulation of the hypothalamo-hypophyseal-gonadal axis. Other endocrine actions, especially a thyrotropic influence, have been postulated. Site and mechanism of action of melatonin, however, are still matters of speculation. In search of a functional cascade of (i) photoperiodic stimuli, (ii) their hormonal messenger melatonin, and (iii) endocrine targets, the pars tuberalis has gained a key position. The recent discovery and characterization of melatonin receptors located in the pars tuberalis of several species support such a functional significance. Earlier results point to a functional connection of the pars tuberalis with the pineal gland: the pars tuberalis is known for a pattern of differentiation distinctly different from other parts of the adenohypophysis. It contains a specific cell population with a morphology typical of peptide secreting cells. Like the hypothalamic nerve endings of the median eminence, they are in close contact with the primary plexus of the portal system. In contrast to secretory cells of the pars distalis, the specific cells of the pars tuberalis do not respond with morphological alterations to functional changes of peripheral endocrine glands. Yet, photoperiodic stimuli obviously influence morphology and functional activity of the pars tuberalis-specific cells. Investigations during recent years have led to the tentative conclusion that the pars tuberalis represents the hypophysial "receptor" for melatonin as the chemical messenger of photoperiodic stimuli. Depending on melatonin secretion pattern and melatonin receptivity, the pars tuberalis seems to modulate at least gonadotropic and thyrotropic activity of the pars distalis via a peptide hormone distributed in the pars distalis by the portal plexus.(ABSTRACT TRUNCATED AT 250 WORDS)


Increased lymphocyte thermogenesis in hyperthyroid patients. Role of Na/K pump function. Evaluation of aerobic/anaerobic metabolism. The role of the Na/K pump for the increased cell energy expenditure in hyperthyroidism was studied by measuring total lymphocyte heat production rate in samples with and without ouabain inhibition of Na/K ATP-ase. In addition, the relative contribution of aerobic processes to lymphocyte thermogenesis was calculated from oxygen consumption measurements. In 12 patients with clinical and laboratory hyperthyroidism total lymphocyte heat production rate was 3.19 +/- 0.21 pW/cell, significantly higher than in 7 patients with subclinical hyperthyroidism (2.14 +/- 0.11 pW/cell) and in 15 euthyroid subjects (2.26 +/- 0.11 pW/cell) (p less than 0.001). The relative decrease in lymphocyte heat production rate after ouabain, giving a quantitative measure of the activity of the Na/K ATP-ase and reflecting the importance of Na/K pump function for the overall rate of lymphocyte metabolism, was not significantly different between the groups: 19.5 +/- 3.6% in hyperthyroid patients, 14.2 +/- 2.3% in subclinical hyperthyroid patients and 17.8 +/- 3.1% in euthyroid subjects. According to the rate of lymphocyte oxygen consumption, aerobic processes represented 58.4 +/- 6.7% of total lymphocyte energy expenditure in hyperthyroid patients, not significantly different from subclinical hyperthyroidism (62.6 +/- 8.4%) or from euthyroidism (66.6 +/- 2.7%). These data do not support the hypothesis of a specific role of the Na/K pump function for the increased cell thermogenesis in hyperthyroidism and indicate a parallel stimulation of aerobic and anaerobic processes by thyroid hormone excess.


Serum estradiol 17-sulphate and lipid peroxides in late pregnancy. Estradiol 17-sulphate is readily converted to 2-OH or 4-OH estradiol 17-sulphate. The latter two strongly antagonize lipid peroxidation, which may play certain roles during pregnancy, such as pregnancy-induced hypertension. Serum estradiol 17-sulphate during mid and late pregnancy was measured using a direct radioimmunoassay without hydrolysis, and the level increased as pregnancy progressed. The levels in the sixth (20-23 weeks) and tenth months (36-39 weeks) of gestation were 1.42 +/- 0.04 nmol/l (mean +/- SD) and 3.42 +/- 1.09 nmol/l, respectively. The maternal venous levels before and at delivery and in the umbilical veins and artery were 3.38 +/- 0.83, 3.48 +/- 1.53, 4.11 +/- 1.40 and 4.30 +/- 1.81 nmol/l, respectively. The latter two values were slightly higher than the former two. Estradiol 17-sulphate in maternal venous blood began to decrease around delivery. Serum lipid peroxides were measured using the method of Yagi. Estradiol 17-sulphate and lipid peroxides showed a simple regression slope (r = -0.548, p less than 0.05) during late pregnancy. These results suggest that estradiol 17-sulphate may be converted to 2-OH or 4-OH estradiol 17-sulphate, which act as lipid peroxide scavengers during pregnancy.


Comparative value of plasma ACTH and beta-endorphin measurement with three different commercial kits for the etiological diagnosis of ACTH-dependent Cushing's syndrome. Recent reports suggest that, contrary to radioimmunoassays (RIA), immunoradiometric assays (IRMA) artifactually decrease plasma ACTH levels in patients with the ectopic ACTH syndrome. Discrepancies between RIA and IRMA results may provide a means of discriminating this entity from Cushing's disease. We have compared the results of these two techniques, together with those of a beta-endorphin assay, in 17 patients with Cushing's disease, 9 with the ectopic ACTH syndrome and 30 controls. ACTH-RIA and ACTH-IRMA levels in patients with Cushing's disease were similar (17.5 +/- 2.5 vs 15.1 +/- 2.8 pmol/l) and were correlated (rs = 0.59, p less than 0.01). ACTH-RIA levels in patients with the ectopic ACTH syndrome were higher than ACTH-IRMA levels (27.3 +/- 2.9 vs 14.5 +/- 2.5, p less than 0.01) and these did not correlate. The ACTH-RIA and ACTH-RIA/ACTH-IRMA ratio levels in patients with the ectopic ACTH syndrome were higher than those of patients with Cushing's disease (p less than 0.01), but they overlapped with these in 27 and 31% of cases respectively. Plasma beta-endorphin level was higher in patients with the ectopic ACTH syndrome than in patients with Cushing's disease (81.9 +/- 19.4 vs 26.4 +/- 5.6 pmol/l, p less than 0.01) and was correlated with ACTH only in patients with Cushing's disease. The overlap in beta-endorphin and beta-endorphin/ACTH-IRMA molar ratio levels between the two groups were 19 and 27% respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Increase in T3 levels during hypocorticism in patients with chronic secondary adrenocortical insufficiency. The clinical and biochemical manifestations of secondary adrenocortical insufficiency are not well defined in the medical literature. This study was designed to determine the clinical and laboratory features suggesting the diagnosis of adrenal insufficiency in 15 chronic ACTH deficiency patients during low and normal cortisol states. Except for fatigue and weakness, the characteristic clinical manifestations of primary adrenal insufficiency occurred rarely. ACTH deficiency did not significantly modify blood glucose, serum calcium, sodium, potassium and differential white blood cell count. However, serum T4 was lower (65 +/- 19 vs 95 +/- 21 nmol/l, p less than 0.001) during cortisol deficiency, while T3 was higher (2.4 +/- 0.67 vs 2.0 +/- 0.60 nmol/l, p less than 0.001). Furthermore, rT3 decreased significantly during hypocorticism (0.27 +/- 0.07 vs 0.18 +/- 0.07 nmol/l, p less than 0.001). The T4/T3 ratio was significantly lower than the normal in 15 out of the 17 episodes of ACTH deficiency (29 +/- 12.5 vs 57 +/- 9.4, p less than 0.0001). We conclude that the increase in T3 and decrease in T4 levels are associated with chronic secondary adrenocortical insufficiency. This laboratory feature could be due, at least in part, to the increased peripheral conversion of T4 to T3 during cortisol deficiency.


Direct effects of bromocriptine on the steroidogenic capability of porcine granulosa cells. The direct effects of bromocriptine on steroidogenesis were examined in cultured porcine granulosa cells. The following observations were made with bromocriptine: (1) It significantly increased the basal or FSH-stimulated secretion of progesterone in cultured porcine granulosa cells at concentrations exceeding 10(-7) mol/l; (2) its inhibitory effect on basal estradiol secretion was demonstrated; (3) it did not influence cell number in cultured porcine granulosa cells; (4) it increased the extracellular accumulation of cAMP in a concentration-dependent manner; and (5) it did not induce a change in cytosolic free Ca2+ concentration. These findings suggest that bromocriptine exerts a direct effect on steroidogenesis in ovarian granulosa cells.


Immunoreactive ACTH in the cerebrospinal fluid of the rat: decrease after mediobasal hypothalamic lesion and hypophysectomy, increase after adrenalectomy. The effect of various anaesthetics and of the manipulations of the hypothalamo-pituitary-adrenocortical system (hypophysectomy, adrenalectomy and lesion of the mediobasal hypothalamus) was studied on immunoreactive-ACTH levels in the plasma and in the cerebrospinal fluid in the rat. The anaesthetics used (Hypnorm, pentobarbital, urethan and Ketanest) were without effect on immunoreactive-ACTH concentration in the cerebrospinal fluid. Immunoreactive-ACTH was significantly decreased after hypophysectomy and elevated after adrenalectomy in both cerebrospinal fluid and plasma. Destruction of the mediobasal hypothalamus resulted in reduced immunoreactive-ACTH content in the cerebrospinal fluid (about 20% of control) in both experiments, whereas immunoreactive-ACTH levels in the plasma of the lesioned rats were lower only in one of the two experiments performed. These data suggest that the main source of the immunoreactive-ACTH in the cerebrospinal fluid of the rat is the hypothalamus; the contribution of the pituitary gland being less than 50% of the radioimmunoassayable ACTH.


The effect of isoflurane on excitatory synaptic transmission in the rat hippocampus. The purpose of this investigation was to study the effect of isoflurane on excitatory synaptic transmission. Rat hippocampal slices maintained in vitro were used as a model. Isoflurane caused a dose-dependent reduction of the excitatory postsynaptic potential (EPSP); 1.5% isoflurane reduced the EPSP by 35 +/- 9% (mean +/- s.d.) and 3% by 57 +/- 11%. Neither spontaneous nor potassium-stimulated efflux of the glutamate analogue D-(3H)aspartate was changed, but the content of D-(3H)aspartate in slices loaded during isoflurane was reduced to 83 +/- 12% of control (P less than 0.05). The intracellularly recorded response to direct application of glutamate increased by 37 +/- 20% during isoflurane (3%) and 50 +/- 5% during halothane (2%). Isoflurane (3%) enhanced the response to the glutamate receptor agonist quisqualate by 44 +/- 19%, whereas the N-methyl-D-aspartate response was unchanged. Isoflurane enhanced the tetanic depression of the population spike. The present results suggest that isoflurane reduces excitatory synaptic transmission by a presynaptic mechanism.


Contraception after thirty-five.

Contraception after thirty-five. Our knowledge about the safety, the incidence of side effects, and the effectiveness of contraceptive methods for women in premenopause, or during the last decade of their reproductive life, has not been a primary interest for research and development in fertility control. The main purpose of trials for the evaluation of new contraceptive methods is to test their effectiveness, and therefore only women below 38 years of age are accepted. Furthermore, when new methods are being tested, only healthy women are accepted, and those having health problems of almost any kind are excluded. Therefore our knowledge about the contraceptive methods in women over 40 years of age is scanty and comes from trials with a long-lasting follow-up on healthy women who can use the method without problems until they are 45 or reach menopause. We have very limited reported information on the use of contraceptives by women who have cardiovascular disease, diabetes, liver problems, etc. Therefore the main body of experience in this review comes from healthy women between 35 and 44. On the other hand, one should remember that these women have a high motivation to use contraceptive methods and therefore the continuation rate in the trials is high. This compensates for the relatively small number of acceptors by giving more women-years in follow-up. Continuation rate and the proper use of methods are directly correlated to increase in age, socioeconomical status, and to education of users. Highly educated women in this age group have very low failure rate with almost any method.


Lynestrenol induced therapeutic amenorrhea: effects of dose reduction on serum sex-hormones and lipids. The effect of reducing the dose of peroral lynestrenol by half on serum sex-hormone, lipid and lipoprotein status was studied in 21 mentally retarded women with therapeutic amenorrhea (TA). They had previously received 5 or 10 mg peroral lynestrenol daily for periods ranging from 32 to 196 months. Dose halving of lynestrenol resulted in an increase in serum total testosterone (T) by 16% (p less than 0.05), sex-hormone binding globulin (SHBG) by 39% (p less than 0.01) and high-density lipoprotein cholesterol (HDL-C) by 28% (p less than 0.001). Both the mean serum total and free concentrations of norethisterone (NET and fNET) decreased by 60% (p less than 0.001). The serum concentrations of 17-beta-estradiol (E2), its free fractions (fE2) and free T (fT) were not significantly altered. Significant correlations were observed between the change in HDL-C and the change in T (r = 0.45, p less than 0.05), between the change in SHBG and the change in T (r = 0.62, p less than 0.01), fT (r = 0.43, p less than 0.05) and E2 (r = 0.51, p less than 0.05). The elevation of HDL-C was probably caused by the reduced serum NET concentrations. This also resulted in an increase in serum SHBG concentration, which is regarded as an indicator of the overall estrogen/androgen ratio.


Analysis of uterine contractility after administration of the non-steroidal anti-inflammatory drug nimesulide. To study the effects of non-steroidal anti-inflammatory drugs (NSAIDs) on uterine contractility in different parts of the uterus and on the direction and velocity of propagation of the activity, intra-uterine pressure (IUP) was measured simultaneously in 10 dysmenorrheic and 5 eumenorrheic patients with two microtransducer catheters at two locations (30 mm apart) before and after taking nimesulide, a newly developed NSAID. The uterus developed higher pressure cycles in the fundus than in the isthmus, in both eumenorrheic and dysmenorrheic conditions. Nimesulide did not affect either the active pressure (AP) or the direction and velocity of propagation of the activity, though it alleviated pain significantly. In dysmenorrheic patients, resting pressure (RP) is at a high level only in the fundus. The velocity of propagation ranged from 12 to 19 mm/s. The mathematical probability of procervical activity (1.0 if all procervical; 0.0 if all profundal), and thus the transport, was 0.59 in eumenorrheic and 0.68 in dysmenorrheic patients, the average for the whole series being 0.65. The luminal content (menstrual blood) moves in the cervical direction much more slowly than would be expected on the basis of simple calculations of velocity (velocity vector) of propagation.


Fetal ponderal index as an instrument for further classification of intra-uterine growth retardation. Perinatal morbidity and mortality are still high in cases of intra-uterine growth retardation. Present screening methods select large risk groups and have a low positive predictive value. Instruments which could be used as indicators for those cases within the risk group needing close fetal monitoring would be valuable. The aim of this prospective study was to evaluate fetal ponderal index with respect to signs of fetal distress and neonatal outcome. By means of a risk scoring system, 73 pregnancies with increased risks of intra-uterine growth retardation were compared with 61 controls. By means of ultrasound, fetal weight was estimated and the fetal femur measured. The fetal ponderal index was calculated by dividing the estimated fetal weight in grams by the third power of the femur length. In the control group, 5/61 showed signs of distress and in the risk group, 43/73. The mean fetal ponderal index of the controls was 8.60 (SD 0.84) and in the risk group 7.72. The groups were compared with each other with respect to signs of distress. The fetuses showing signs of distress had a mean FePI of 7.45 (p less than 0.001). Those (30/73) in the risk group not showing signs of fetal distress had a mean value of 8.14 and differed significantly (p less than 0.001) from the distress group. Fetal ponderal index would seem to be a valuable adjunct for the differentiation of the more susceptible fetuses in an intra-uterine growth retardation risk group.


An open study comparing two regimens of gemeprost for the termination of pregnancy in the second trimester. Two regimens of the prostaglandin E1 analogue, gemeprost, were compared in an open trial for termination of pregnancy between 12 and 18 weeks. Fifty women received 5 x 1 mg of gemeprost every 3 h and in another 50 cases, 4 x 1 mg of gemeprost was administered every 6 h. Although the median abortion interval was slightly shorter in the 3-hourly group (15.9 h vs. 16.9 h; p = 0.5), the cumulative abortion rates at 24 h were similar (88% vs. 82%; p less than 0.5). In women who aborted within the first 24 h, significantly fewer pessaries (p less than 0.01) were required to induce abortion in the 6-h treatment group (median 3, range 1-4) than the 3-h group (median 5, 2-5). Parous women in both treatment groups required fewer pessaries to induce abortion than did nulliparous women (not significant; p = 0.5). Significantly (p less than 0.01) fewer pessaries were required to induce abortion in the 6-h gemeprost group. The were no significant differences between the groups regarding incidences of diarrhea, vomiting, or the request for analgesia. These results suggest that in many women the number of pessaries used to induce mid-trimester abortion could be reduced by lengthening the interval between insertion of pessaries within the first 24 h, without loss of clinical efficacy.


Prolactin and amniotic fluid electrolytes. Prolactin (PRL) levels and Na+, K+, Cl-, Ca++ concentrations in maternal serum and amniotic fluid from 64 women in normal term pregnancy were measured by immunoenzymetric assay and flame photometry. The mean amniotic fluid PRL concentration was 597.7 (SE 31.5) ng/ml and the mean amniotic fluid Na+, K+, Cl- and Ca++ levels were 125.6 (SE 0.9) mmol/l, 4.5 (SE 0.1) mmol/l, 109.3 (SE 1.3) mmol/l and 2.0 (SE 7.5 E-02) mmol/l, respectively. There was no correlation between PRL levels in maternal serum and amniotic fluid, and the electrolyte concentrations in amniotic fluid. A close correlation was found between the concentrations of Na+ and Cl- in maternal serum and amniotic fluid. Thus, even though PRL may participate in the regulation of electrolytes in the amniotic fluid compartments, our findings provide indirect evidence for the existence of other regulatory mechanisms.


Induction of labor by vaginal prostaglandin E2. A randomized study comparing pessaries with vaginal tablets. A prospective randomized study of 267 pregnant women was undertaken to compare the efficacy of a pharmacy-prepared 3-mg prostaglandin E2 (PGE2) vaginal suppository with a 3-mg PGE2 vaginal tablet for induction of labor and cervical ripening. No statistically significant difference in success frequency was found between the two groups, either on the first day (72% and 74%, respectively; p greater than 0.05) or on the second day (89% in both groups). There was an equal proportion of women requiring oxytocin augmentation in the two groups, but the slower releasing properties of the vaginal tablet were reflected in a longer mean induction--delivery interval of about 4 h for this group. In both the pessary and the vaginal tablet groups, women who had not gone into labor on the first day showed a statistically significant increment in the Bishop score on the morning of the second day. The frequency of cesarean section was the same in both groups, but instrumental deliveries were more frequent in the vaginal tablet group. It is concluded that PGE2 vaginal tablets--a chemically stable alternative to pharmacy prepared pessaries--appear to be effective as regards cervical ripening as well as for labor induction.


Reproductive patterns among Swedish women born 1936-1960. An analysis was made of a cross-sectional survey of 4,299 women born in Sweden in the period 1936-1960, regarding their reproductive performance. Fecundability was only 12% lower for women in the 30-39 year age interval than for women up to 29 years of age. Cumulative pregnancy rates for individual years fell in the range 93-96%. No difference in fecundability could be discerned between women born in 1936-45 vis-à-vis 1946-60. Primary and secondary infertility were noted for 6.6% and 11.0% respectively of the entire population surveyed. Primary infertility, more prevalent among those born in 1946-60, was found to be related to a higher level of education, an urban life style and low age at first coitus. The risk of suffering a spontaneous abortion increased with advancing maternal age.


Semen quality of smoking and non-smoking men in infertile couples in a Swedish population. A study of semen quality, using conventional semen analysis, was undertaken. 186 male smokers were compared with 164 non-smoking men undergoing infertility investigation. The cigarette smokers were subdivided into three groups according to the number of cigarettes they smoked: less than 10 (n = 27), 10-19 (n = 84) and greater than 19 cigarettes per day (n = 75). No statistically significant effect of cigarette smoking on sperm density, motility or morphologic features of sperm was detected. Nor was any significant difference in sperm quality, except for semen volume and total sperm count, disclosed between men in the different smoking categories or between heavy smokers and non-smoking men. Thus, the present study does not corroborate reports of detrimental effects of cigarette smoking alone on sperm concentration, motility or morphology on the bases of this population of Swedish men in infertile couples.


'Hit and run' oncogenesis by human papillomavirus type 18 DNA. Transfection of an immortalized cell line (AE), derived from Syrian hamster embryo cells, with human papillomavirus type 18 (HPV 18) DNA induced morphological transformation and these transformed cells were tumorigenic in nude mice. Southern blot analysis revealed that the transfected viral DNA was retained in all the cell lines tested, however, all these transformed cells contained only less than one copy per cell of viral genome. Eleven cloned cell lines were established from a tumor cell line obtained after explantation of a tumor into a nude mouse. Two lines revealed no viral sequences by both Southern blot hybridization and polymerase chain reaction, whereas the nine others contained the remaining viral sequences. These results are highly suggestive of a 'hit and run' oncogenesis by this virus.


Laparoscopic hysterectomy. Initial experience. Laparoscopic hysterectomy is now being performed in our department in cases where no malignancy is suspected. This study presents the first 10 cases. Indications for hysterectomy were myomas, meno- and metrorrhagia resistant to medical and hysteroscopic treatment, and patients with pain and suspicion of having adenomyosis. No complications have been encountered during the laparoscopic operations, but one patient had a second laparoscopy on the first postoperative day due to postoperative bleeding. Another patient had a postoperative infection leading to a compression of the ureter. This report demonstrates that laparoscopic hysterectomy is a valuable addition to the new procedures in 'minimal invasive surgery', but only after long and appropriate training.


Consumer's attitude to hysterectomy. The experience of 678 women. A total of 678 women were interviewed about symptoms before as well as advantages and disadvantages after abdominal hysterectomy. Medical complications, side effects, days of hospitalization and some possible differences between supravaginal hysterectomy (SVH) and total hysterectomy (TH) were also investigated. TH had been performed in 79% and SVH in 21% of the women. Leiomyoma was the most frequent diagnosis (79%). Heavy menstrual bleeding and dysmenorrhea were the dominant symptoms before surgery. After the operation, 71% of the women had no subjective gynecological complaints. Relief from heavy bleeding and pain was considered the major advantage and as many as 29% of the women experienced no disadvantage at all with the operation. Regarding sexual life, 39% experienced intercourse as much improved or better and 40% as unchanged. An intraabdominal abscess was more often found postoperatively in patients operated with TH than with SVH but with this exception no objective finding favoured one method more than the other. When new, alternative treatments such as endometrial ablation and GnRH analogues are introduced and promoted they should be subjected to careful evaluation and compared with the therapeutic efficacy of hysterectomy.


Rupture of rudimentary horn pregnancy.

Rupture of rudimentary horn pregnancy. A rare case of rupture of an noncommunicating rudimentary horn pregnancy is reported. The pregnancy proceeded to 25 gestational weeks, when the rudimentary horn ruptured. The patient had signs and symptoms of massive hemoperitoneum. An emergency exploratory laparotomy revealed rupture of the gravid rudimentary horn and the fetus was lying free in the peritoneal cavity, was delivered breech and died 2 hours later. This reproductive complication was treated successfully by prompt exploratory laparotomy. Some of the factors associated with pregnancy in a rudimentary horn are discussed.


Indomethacin treatment for polyhydramnios. Effective but potentially dangerous? A case of acute polyhydramnios for unknown reason was successfully treated with orally administrated indomethacin. The need for close supervision of the pregnancy, because of potential risk for oligohydramnios, is described.


Amelanotic malignant melanoma arising in an ovarian dermoid cyst. Malignant transformation occurs in about 2% only of ovarian dermoid cysts. The clinical and histopathological findings in a rare case of primary ovarian malignant melanoma of amelanotic type are presented.


Combined ovarian vein catheterization with ovarian stimulation in the diagnosis of androgen overproduction. A 28-year-old woman was evaluated for late onset secondary amenorrhea, progressive hirsutism and an elevated serum testosterone concentration. Her serum cortisol, androstenedione, dehydroepiandrosterone sulfate and 17-hydroxyprogesterone levels were normal. Bilateral ovarian and adrenal vein catheterization demonstrated mild elevated testosterone and androstenedione levels in the right ovarian vein. Fifteen minutes after administering the intravenous injection of 5,000 IU human chorionic gonadotropin, there was a six and a half to sevenfold increase in the level of these two hormones in the right ovarian vein with no significant change in hormone levels from other sources. Based on the ovarian peripheral vein gradients obtained during venography following ovarian stimulation, the diagnosis of right ovarian hyperthecosis was made. This diagnosis could not have been reached without the combination of selective ovarian vein catheterization and ovarian stimulation. We recommend that this combined test, which may provide additional information on the source of the androgens in women with hyperandrogens, be performed in selected cases, when a virilizing tumor is suspected.


Adenosine receptor-induced cAMP changes in D384 astrocytoma cells and the effect of bradykinin thereon. In human D384 astrocytoma cells, cyclic AMP accumulation can be conveniently studied after labelling of the adenosine triphosphate pool (15 fmol cell-1) with [3H]adenine. In this study, adenosine had a biphasic effect on cyclic AMP accumulation, which was scarcely altered by blocking adenosine uptake and metabolism. Low concentrations of adenosine led to an inhibition of cyclic AMP accumulation, and higher concentrations led to stimulation. No effect of adenosine on cyclic AMP was observed unless phosphodiesterase was inhibited by rolipram. The A1 receptor antagonist DPCPX attenuated the inhibitory phase of adenosine response, and enhanced the cyclic AMP accumulation induced by adenosine analogues. The cyclic AMP accumulation was stimulated by NECA greater than ADO greater than CGS 21680 greater than CV 1808 greater than CPA greater than or equal to CHA, indicating mediation by A2 receptors. The stimulatory effect of NECA was much more effectively blocked by the combined A1 and A2 receptor antagonist CGS 15943 (KB 4 nmol l-1) than by the A1 antagonist DPCPX (KB 110 nmol l-1). Treatment of the cells with pertussis toxin (0.2 microgram ml-1 for 2.5 h) potentiated the cyclic AMP response to adenosine analogues significantly. The cyclic AMP response to NECA was enhanced by the protein kinase C activator phorbol dibutyrate even after pertussis toxin treatment. By contrast, nanomolar concentrations of bradykinin, which increases Ca(2+)-levels and protein kinase C activity in D384 cells, reduced NECA-induced cyclic AMP accumulation in control and pertussis toxin-treated cells. Thus, D384 cells possess both A1 and A2 adenosine receptors influencing cyclic AMP in opposite directions.(ABSTRACT TRUNCATED AT 250 WORDS)


Growth inhibition of human hepatoma cells (HepG2) by polyunsaturated fatty acids. Protection by albumin and vitamin E. Albumin carries fatty acids and has also been suggested to act as an antioxidant. In the present work, polyunsaturated fatty acids (linoleic, arachidonic, eicosapentaenoic and docosahexaenoic acids)--but not palmitic and oleic acid--inhibited growth of human hepatoma cells in low albumin concentration (0.5%). Growth inhibition by polyunsaturated fatty acids was prevented by albumin in a dose-related manner in the range 0.7-5.0%. Albumin also protected against growth inhibition following catabolism (by lipoprotein lipase) of very low density lipoproteins. Vitamin E strongly counteracted the inhibitory effect of polyunsaturated fatty acids. Vitamin E and albumin appeared to have additive effects in protecting against growth inhibition by polyunsaturated fatty acids. Indomethacin did not greatly modify the polyunsaturated fatty acids effect. Growth inhibition by polyunsaturated fatty acids, as well as the level of thiobarbituric acid reacting substances (a measure of lipid peroxidation) in growth media, increased with increasing number of fatty acids double bonds. Vitamin E and albumin prevented both thiobarbituric acid reacting substances formation and growth inhibition by polyunsaturated fatty acids. The results suggest that the concentrations of albumin and vitamin E in the incubation medium are essential when studying polyunsaturated fatty acids effects on cell growth.


Neural symptoms induced by tricyclic antidepressants: phenomenology and pathophysiology. The authors review the literature describing the neural symptoms induced by tricyclic antidepressants, especially tremor, seizures, akathisia, myoclonus, dyskinesia and delirium. Sedation, modifications of sleep, memory and appetite are also described. Tremor and myoclonus are the most frequent drug-induced neural symptoms. Delirium is most often caused by high-dosage treatments. The pathophysiology of akathisia and dyskinesia raises important questions concerning the mode of action of antidepressants.


[Prevention of thrombogenesis in the extracorporeal circuit of hemodialysis with a low molecular heparin: standardization of the dosage with a better hemorrhagic risk/effectiveness ratio]. To test the efficacy and safety of a low molecular. Weight heparin (LMWH)--Fraxiparine, for hemodialysis (HD) anticoagulation, compared with conventional heparin (H) or serum lavage without other anticoagulation (L). Prospective controlled study. Twenty-nine consecutive patients referred for dialysis in a tertiary care hospital were divided in 3 groups A, B and C, each group A and B patient submitted to 2 dialysis, AI and AII, BI and BII. Group A--n = 10, no bleeding risk, single needle technique, blood flow (Qb) less than 200 ml/min. HD-AI used LMWH 10,000 U pre-HD, HD-AII used H for an ACT 1.5 to 2 times baseline; group B--n = 10, high bleeding risk, double needle dialysis, Qb--200 to 300 ml/min. HD-BI used LMWH 5000 U pre-HD, and HD-BII used only L; Group C--n = 9, no bleeding risk, Qb less than 200 ml/min, all received LMWH 5000 U pre-HD. A semiquantitative screening was done in each dialysis for the presence of dialyser or venous chamber clots, APTT and Anti Xa activity were measured every 30 min., as well as pre and post-dialysis Hb, Htc, and platelets. APTT didn't rise significantly during HD with LMWH in contrast with the AII group with H (32.2 +/- 7.1 vs 63 +/- 25.8, p less than 0.05). The APTT levels in all dialysis with LMWH were identical to BII dialysis With L. Anti xa activity had an early peak at 30 to 60 min. With LMWH (0.62 +/- 0.45 em AI) and a late one at 180 min with H (0.39 +/- 0.2). There was no significant differences between pre and post-dialysis corrected platelet counts, but the lavage group showed the greater decrements (-20% +/- 24). In all the 49 dialysis we had 5 cases of complete clotting of the blood circuit, all of them in the lavage group C. No patients with high risk of hemorrhage had any bleeding increment. LMWH prevents clotting as effectively as H, in low doses of 5000 anti Xa units it doesn't interfere with PTT and is far more effective than HD with serum lavage in patients with bleeding risk and/or low blood flow in the dialysis circuit.